Substituted RIG-I agonists: compositions and methods thereof

ABSTRACT

This invention provides compositions, compounds, and uses thereof, wherein said compounds comprise a single strand oligonucleotide that may form a short oligonucleotide hairpin or stem loop molecule with self complementary base pairing of less than 12 base pairs that bind to RIG-I and activate the RIG-I pathway.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a National Stage application of International Patent Application No. PCT/US2019/027423, filed Apr. 15, 2019, which claims the benefit of priority to U.S. Provisional Patent Application No. 62/660,568, filed Apr. 20, 2018, the contents of each of which are hereby incorporated by reference in their entireties.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jun. 22, 2021, is named 24588USPCT-SEQLIST-22JUN2021.txt and is 2320 Kb in size.

BACKGROUND OF THE INVENTION

Retinoic acid-inducible gene 1 (RIG-I) plays a crucial role in promoting the release of type I and type III interferons to fortify host anti-viral immunity (Wu et al., 2015, Virology 482: 181-188). The RIG-I pathway has been identified as an important innate immune sensing pathway of cytosolic 5′-triphosphate (5′ppp) dsRNA that drives the production of a variety of cytokines, including interferon β. In response to viral 5′ppp cytosolic double stranded RNA, RIG-I undergoes activation and recruits mitochondrial antiviral signaling protein (MAVS). Moreover, transcriptome analysis reveals a RIG-I-related signature covering the canonical pathway categories “IFN signaling”, “activation of IRFs by cytosolic pattern recognition receptors (PRRs)”, “TNFR2 signaling” and “antigen presentation” indicating that RIG-I bridges the innate and adaptive immune system (Goulet et al., 2013, PLoS Pathogens). Intriguingly, RIG-I-induced immunogenic tumor cell death triggers adaptive immunity, engaging dendritic cells and T-cells to kill tumors in vivo providing a second innate/adaptive immune system loop (Duewell et al., Cell death and differentiation 2014: 21(12) 1825-1837). Of note, RIG-I-induced apoptosis is restricted to tumor cells only (Duewell et al., 2014, 21(12) 1825-1837).

Recent studies identified key structural features for optimal RIG-I ligands. Short length, double-strandedness, 5′-triphosphorylation and blunt end base pairing characterize the prototypic RNA-based RIG-I agonist (Schlee et al., 2009, Immunity 31:25-34; Pichlmair et al., Science 2006, 314: 997-1001; Schlee, Immunobiology, 2013; 218(11), 1322-1335). Moreover, circular structures (Chen et al. Mol Cell 67 (2), 228-238.e5. 2017) and bent/KINK RNAs (Lee et al., 2016, Nucleic Acids Res, 44(17), 8407-8416) constitute another recently identified group of RIG-I ligands that do not require a triphosphate moiety. Nabet et al. (Cell 2017 Jul. 13; 170(2):352-366.e13. doi: 10.1016/j.cell.2017.06.031) reported that an unshielded endogenous RNA can activate RIG-I in tumor cells promoting aggressive features of cancer. Recent findings indicate also that endogenous small non-coding RNAs leaking to the cytoplasm can activate RIG-I during ionizing radiation therapy (Ranoa et al., Oncotarget 2016; 7(18), 26496-26515). In addition, a hetero-trimeric complex of RIG-I/RNA polymerase III/serine-arginine-rich splicing factor 1 facilitates RIG-I activation in response to delocalized, cytosolic DNA via a 5′ppp RNA intermediate (Ablasser et al., Nat Immunol 2009; 10(10), 1065-1072; Xue et al., PLoS One 2015; 10920, E0115354).

Structural and functional analysis of RIG-I reveals that single amino acids and a lysine-rich patch located at the C-terminal domain (CTD) of RIG-I sense the structural properties of RNAs (Wang et al., Nat Struc Mol Biol 2010; 17(7), 781-787). Remarkably, a typical eukaryotic 2′-O-methylation pattern along with 7-methyl guanosine capping of the 5′-triphosphate group of RNAs prevent binding to RIG-I, thus distinguishing host from pathogenic non-self RNA. Specifically, modifications at the 5′ end decrease RNA affinity and production of pro-inflammatory cytokines (Schuberth-Wagner et al., 2015, Immunity 43, 41-51; Devarkar et al., PNAS 2016; 113(3), 596-601). Modified RNAs containing modified nucleotides m6A, Ψ, mΨ, 2FdU, 2FdC, 5mC, 5moC, and 5hmC appear to lack stimulatory activity (Durbin et al., mBio 7(5):e00833-16. doi: 10.1128/mBio.00833-16). Moreover, illegitimate RIG-I activation by endogenous RNA is controlled by fast ATPase turnover which leads to dissociation of the RIG-I/RNA complex (Louber et al., BMC Biology 2015; 13:54). Interestingly, RIG-I mutations that correlate with a decreased ATPase activity appear to be constitutively active potentially due to signals from host RNA (Fitzgerald et al., 2017, Nucleic Acids Res, 45(3), 1442-1454).

Improvement in the understanding of how RIG-I recognizes RNA and utilizes ATP has resulted from structural studies of RIG-I truncations in mouse, human, and duck (Civril et al., 2011, EMBO Rep 12:1127-1134; Jiang et al., 2011, Nature 479:423-427; Kowalinski et al., 2011, Cell 147:423-435; Luo et al., 2011, Cell 147:409-422). However, in the only RIG-I structure with caspase activation and recruitment domains (CARD) present, the protein was in an inactive, apo-state, and lacked the CTD. This leads to questions regarding the role of both RNA and ATP in the innate immune response derived from RIG-I activation, and the relative positions of the CTD and CARDs in the active RIG-I conformation. Interestingly, the RIG-I CTD capped the 5′ end of the RNA, regardless of the length of the bound duplex. The preference for the end of the duplex RNA in these structures was also independent of a 5′ppp. Recent publications, including those mentioned herein, have demonstrated that activation of the innate immune system through RIG-I leads to potent anti-tumor immunity. This new immunotherapeutic mechanism may be synergized with checkpoint inhibitors such as an anti-PD1 antibody to achieve greater anti-tumor responses. See also Elion et al., 2018, Cancer Research (DOI: 10.1158/0008-5472.CAN-18-0730).

Hence, there still remains a need for understanding RIG-I functionality and establishing RIG-I selectivity, boosting or abrogating RIG-I-driven immunity.

SUMMARY OF THE INVENTION

The present invention provides a composition comprising a nucleic acid capable of activating RIG-I. This invention provides compounds, and uses thereof, wherein said compounds comprise a short single strand oligonucleotide that optionally forms an oligonucleotide hairpin or stem loop molecule with self complementary base pairing of less than 12 base pairs that bind to RIG-I and activate the RIG-I pathway. An embodiment of this invention is realized when the short single strand oligonucleotide does not form an oligonucleotide hairpin or stem loop molecule.

An embodiment of this invention is realized when the short single strand oligonucleotide forms an oligonucleotide hairpin or stem loop molecule. An embodiment of this invention is realized when the short oligonucleotide hairpin molecule optionally comprises a 5′-terminus group selected from OH, diphosphate (pp) group and triphosphate (ppp) group. Another embodiment of this invention is realized when the short oligonucleotide hairpin molecule optionally comprises a 5′-terminus selected from functional groups that mimic the interactions of the triphosphate and diphosphate groups, sometimes referred to as 5′ terminus pp mimic or ppp mimic. An embodiment of this invention is realized when in the absence of a 5′-terminus phosphate group the short oligonucleotide molecule comprises a 5′-terminus that is a 5′-OH. Another embodiment of this invention is realized when the short oligonucleotide hairpin molecule comprises a 5′ terminus pp or 5′ terminus pp mimic. Another embodiment of this invention is realized when the short oligonucleotide hairpin molecule comprises a 5′-terminus ppp, or a 5′-terminus ppp mimic.

An embodiment of this invention is realized when the short oligonucleotide molecule comprises a 5′-terminus ppp. Still another sub-embodiment of this invention is realized when the short oligonucleotide molecule comprises a 5′-terminus ppp mimic. In yet another embodiment the short oligonucleotide hairpin molecule comprises a 5′-terminus group selected from diphosphate and triphosphate, a self complementary base pairing region of less than 12 base pairs, and a blunt end. In another embodiment, the short oligonucleotide hairpin molecule comprises a 5′-terminus mimic group selected from diphosphate and triphosphate, a self complementary base pairing region of less than 12 base pairs, and a blunt end. In another embodiment, the short oligonucleotide hairpin molecule comprises a 5′-terminus mimic group selected from diphosphate and triphosphate, a self complementary base pairing region of less than 12 base pairs, a blunt end, and a non-base pairing stem loop. A subembodiment of this aspect of the invention is realized when the looped end is a non-base pairing tetraloop consisting of four nucleotides. Another subembodiment of this aspect of the invention is realized when a blunt end is at the 5′terminus end.

Still in another embodiment, the invention is directed to short oligonucleotide hairpin or stem loop molecules having a self complementary base pairing region of less than 12 base pairs, a blunt end, and a non-base pairing tetraloop, further comprising a modified phosphodiester backbone. A sub-embodiment of this aspect of the invention is realized when the modified phosphodiester backbone comprise at least one modified phosphate group comprising a phosphorothioate.

In another embodiment of this aspect of the invention, the short oligonucleotide hairpin molecules comprise at least one modified nucleotide base (nucleobase).

In yet another embodiment, disclosed is a method for inducing type I interferon production in a cell, or a method for treating a disease or disorder in a subject in need thereof by inducing type I interferon production in a cell of the subject. In another embodiment, use of one or more short oligonucleotide molecules in the potential treatment or prevention of RIG-I pathway-associated diseases or disorders is disclosed, wherein the molecule comprises a self-complementary base pairing region of less than 12 base pairs, a blunt end, and non base pairing tetraloop. In a sub-embodiment of this aspect of the invention, the disease or disorder is selected from the group consisting of a bacterial infection, a viral infection, a parasitic infection, cancer, an autoimmune disease, an inflammatory disorder, and a respiratory disorder.

In still another embodiment, compositions comprising one or more of short oligonucleotide molecules are disclosed. In another embodiment, a pharmaceutical composition comprising one or more of the short oligonucleotide hairpin molecules capable of inducing interferon production and a pharmaceutically acceptable carrier, wherein the molecule comprises a self complementary base pairing region of less than 12 base pairs, a blunt end and a non base pairing tetraloop. In a sub-embodiment of this aspect of the invention, the 5′-terminus of the short oligonucleotide hairpin molecules comprise at least one of the group consisting of 5′-OH, 5′-triphosphate, 5′-diphosphate, 5′-diphosphate mimic, and 5′-triphosphate mimic. In a sub-embodiment of this aspect of the invention, the pharmaceutical composition further comprises at least one agent selected from an immunostimulatory agent, an antigen, an anti-viral agent, an anti-bacterial agent, an anti-tumor agent, retinoic acid, IFN-α, and IFN-β.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to novel oligonucleotides and analogues thereof and their use to bind and activate the RIG-I protein.

Embodiments of the disclosure include compounds of general Formula Ia and Formula I as well as synthesis and isolation of compounds of general Formula Ia and Formula I. Uses of compounds of general Formula Ia and Formula I are also disclosed. Other embodiments, aspects and features of the present disclosure are either further described in or will be apparent from the ensuing description, examples and appended claims.

The term “oligonucleotide” in the context of the present application encompasses compounds comprising a plurality, e.g., at least 4 nucleotide or nucleotide analogue building blocks, preferably, 6-100, e.g., 19-40, 19-30, and 19-25 building blocks. The nucleotide or nucleotide analogue building blocks may comprise nucleoside or nucleoside analogue subunits connected by inter-subunit linkages. The nucleoside subunits include deoxyribonucleoside subunits, ribonucleoside subunits and/or analogues thereof, particularly sugar- and/or nucleobase-modified nucleoside analogues. Further, the oligonucleotides may comprise non-nucleotidic building blocks and/or further terminal and/or side-chain modifications.

Examples of sugar-modified subunits are realized when R₁, R₂ R₃, and/or R₄ of a ribonucleoside subunit is replaced, for example, by a group as defined below. In other sugar-modified subunits, the ribose may be substituted, e.g., by another sugar, for example a pentose such as arabinose. This sugar modification may be combined with 2′-OH modifications as described above, such as in 2′-fluoroarabinonucleoside subunits. Still other sugar-modified subunits include locked nucleosides (LNA). A “locked nucleoside” is a nucleoside in which the robose moiety is modified with an extra bridge connecting a 2′ oxygen and a 4′ carbon. Still other nucleobase-modified nucleosidic building blocks, a non-standard, e.g., non-naturally occurring nucleobase, is used instead of a standard nucleobase. Examples of non-standard or modified nucleobases are uracils or cytosines modified at the 5-position, e.g., 5-(2-amino)propyl uracil or 5-bromouracil; hypoxanthine; 2,6-diaminopurine; adenines or guanines modified at the 8-position, e.g. 8-bromoguanine; deazanucleosides, e.g. 7-deazaguanine or 7-deazaadenine; or O- and N-alkylated nucleobases, e.g. N⁶-methyladenine, or N⁶,N⁶-dimethyladenine. Further suitable nucleobase analogues may be selected from universal nucleobase analogues such as 5-nitroindole or inosine.

As used herein, the term “backbone” refers to the linkage between the subunits of the oligonucleotide. A “modified phosphodiester backbone” occurs when the inter-subunit linkage between subunits is be a phosphodiester linkage or a modified linkage, e.g. a phosphorothioate, phosphorodithioate, methylphosphonate, phosphoramidate, boranophosphate, and the like or another modified linkage described herein or known to a skilled person in the art.

As used herein, a “single stranded oligonucleotide” refers to an oligonucleotide in which the bases are not complementary to each other. A “hairpin” oligonucleotide structure refers to a single stranded oligonucleotide, wherein a portion of the oligonucleotide chain loops around and self complementary base pairing results in a portion of the oligonucleotide which is effectively double stranded.

As used herein, a “double stranded oligonucleotide,” refers to an oligonucleotide of two strands, which is formed from complementary base pairsA double-stranded oligonucleotide as used herein can have terminal overhangs on either end or both ends. An oligonucleotide section is fully double-stranded when the two stretches of nucleic acid forming the section have the same length and have sequences which are 100% complementary to each other. As established in the art, two nucleotides are said to be complementary to each other if they can form a base pair, either a Waston-Crick base pair (A-U, G-C) or a wobble base pair (U-G, U-A, I-A, I-U, I-C).

As used herein, the term “blunt end” as used in a double stranded oligonucleotide refers to the 3′ and 5′ ends of the oligonucleotide of the invention, wherein the oligonucleoide terminates in a base pair. Oligonucleotides which do not have a blunt end are referred to as having an overhang, wherein one strand has a longer oligonucleotide chain than the other strand. A single stranded oligonucleotide may have a single blunt end or an overhang.

The oligonucleotide may be selected from deoxyribonucleotides, ribonucleotides and oligonucleotide analogues. The deoxyribonucleotides, ribonucleotides and/or oligonucleotide analogues may be chemically modified at the nucleoside and/or ribose subunit of the analogue, deoxyribonucleotide, ribonucleotide and/or oligonucleotide. Analogues of desoxyribonucleotides or ribonucleotides may comprise at least one desoxyribonucleoside or ribonucleoside subunit and at least one modified nucleosidic subunit and/or at least one modified inter-subunit linkage, e.g. as described above. Oligonucleotide analogues may also consist in their entirety of modified nucleosidic subunits.

The oligonucleotides of the present invention comprise short single strand oligonucleotides that optionally may fold to form a hairpin consisting of self complementary base pairings and an unpaired tetraloop. The self complementary base pairing hairpin structure is formed when intra-molecular base pairing forms a double helix that ends in an unpaired tetraloop opposite the 5′ terminus end of the single strand. An embodiment of this aspect of the invention is realized when the self complementary base pairing is less than 12 base pairs and the unpaired loop is a tetraloop of four nucleotides. The hairpin molecules may be blunt ended or comprise at least one overhang, e.g., a 5′- or 3′-overhang. The hairpin molecules may optionally comprise a 5′ OH group, 5′-terminus phosphate group or 5′-terminus phosphate mimic group.

The oligonucleotides of the present invention comprise single strands of a length of at least 24 nucleotides. In another embodiment, the single strand has a length of 24 nucleotides.

The oligonucleotide may comprise further terminal and/or side-chain modifications, e.g., cell specific targeting entities covalently attached thereto. Those entities may promote cellular or cell-specific uptake and include, for example lipids, small molecules, cholesterol, folate, vitamins, hormones, peptides, oligosaccharides and analogues thereof. Targeting entities may, e.g., be attached to modified nucleobases or non-nucleotidic building blocks by methods known to the skilled person.

Still in another embodiment, modifications establish and/or enhance the selectivity of the oligonucleotide towards a given target. In a particularly preferred embodiment the RIG-I selectivity of the oligonucleotide is enhanced. Methods to determine the RIG-I selectivity of a given oligonucleotide are described herein in detail (cf. Examples) and/or are known to the person skilled in the art.

Yet in another embodiment, the chemical modifications maintain or enhance the chemical stability of the oligonucleotide. A person skilled in the art knows methods for determining the chemical stability of a given oligonucleotide. Such methods are also described, e.g., in the Examples.

Chemical modifications of the oligonucleotide include those to the ribonucleoside subunit, phosphodiester backbone, and nucleobase. Non-limiting examples of such modifications are independently selected from the group comprising halogenation, (e.g., 2′-F-halogenation), 2′-O-alkylation (e.g., 2′-O-methylation), and/or phosphorothioate modifications of internucleotide linkages. Particularly, 2′-F-halogenation and phosphorothioate modifications may increase the stability of the oligonucleotide, while 2′-O-methylation may establish or increase RIG-I selectivity of the oligonucleotide. 2′-O-methylations may also be able to modify the immunogenicity of RNA.

The identification patterns of a given oligonucleotide depend on the sequence and the length of an oligonucleotide and can be determined for each given oligonucleotide. A person skilled in the art is well aware how to carry out this determination. As explained herein, such methods for determining RIG-I selectivity and/or stability of a given oligonucleotide are described in detail in the present application.

The present invention provides modified short oligonucleotide molecules of general Formula Ia:

or pharmaceutically acceptable salts thereof, wherein: each X is independently selected from the group consisting of O, S, —CH₂—, and —NH—; each Y is independently selected from the group consisting of —OH, —SH, and

each V is independently O, or S; K is H, or

each W is independently N, —CH—, or —C(F)—; each Z is independently N, or —CH—; each R₁ is independently selected from the group consisting of H, halogen, —OH, —OC₁₋₆ alkyl, C₁₋₆ alkyl, —OC₁₋₃ haloalkyl, —OC₂₋₆ alkenyl, —OC₂₋₆ alkynyl, —NH₂, and —OCH₂C₅₋₆ heteroarylCH₂R_(y), said alkyl, alkenyl, alkynyl and heteroaryl optionally substituted with 1 to 3 groups of R_(b); each R₂ is independently selected from the group consisting of H, halogen, —OH, —OC₁₋₆ alkyl, C₁₋₆ alkyl, —OC₁₋₃ haloalkyl; or one or more adjacent R₁ and R₂ can combine to form ═CH₂; each R₃ is independently H, halogen, C₁₋₆ alkyl, or C₂₋₆ alkynyl; or R₁ and R₃ on a sugar moiety can be linked to form —CH₂O, provided R₂ is hydrogen; R₄ is selected from the group consisting of —CH₂OH,

R₅ is selected from the group consisting of H, C₁₋₆ alkyl, —(CH₂CH₂O)_(p)CH₂CH₃, and —CH₂CH(OH)CH₂(O(CH₂)₂)_(n)CH₂NHC(O)R_(z); each R₆ is independently selected from H and NH₂; R_(b) is selected from C₁₋₆ alkyl, —OC₁₋₆ alkyl, halo, OH, CN, NO, NH₂, NHC₁₋₆alkyl, N(C₁₋₆alkyl)₂, SO₃H, SO₄, PO₄, C₁₋₃ haloalkyl, C₁₋₃ haloalkoxy, carboxyl, oxo, and S(O)_(g)alkyl; R is H or —P(O)(OH)_(2′); Q is independently selected from the group consisting of O, CH₂, and NH; R_(s) is NH₂ or —C(O)—; R_(x) is hydrogen or —C≡CH; R_(y) is —CH₂(O(CH₂)₂)_(n)(CH₂)_(k)NHC(O)—R_(z), or —(CH₂OCH₂)_(n)CH₂C(O)N((CH₂)_(k)NHC(O)R_(z))₂; R_(z) is cholesterol; g is 1, 2, or 3; k and n are independently selected from 1, 2, 3, 4, 5, and 6; p is an integer from 1 to 50.

An embodiment of the invention of Formula Ia is represented by Formula I:

or pharmaceutically acceptable salts thereof, wherein: each X is independently selected from the group consisting of O, S, —CH₂—, and —NH—; each Y is independently selected from the group consisting of —OH, —SH, and

each V is independently O, or S; K is H, or

each W is independently N, —CH—, or —C(F)—; each Z is independently N, or —CH—; each R₁ is independently selected from the group consisting of H, halogen, —OH, —OC₁₋₆ alkyl, C₁₋₆ alkyl, —OC₁₋₃ haloalkyl, —OC₂₋₆ alkenyl, —OC₂₋₆ alkynyl, —NH₂, and —OCH₂C₅₋₆ heteroarylCH₂R_(y), said alkyl, alkenyl, alkynyl and heteroaryl optionally substituted with 1 to 3 groups of R_(b); each R₂ is independently selected from the group consisting of H, halogen, —OH, —OC₁₋₆ alkyl, C₁₋₆ alkyl, —OC₁₋₃ haloalkyl; or one or more adjacent R₁ and R₂ can combine to form ═CH₂; each R₃ is independently H, halogen, C₁₋₆ alkyl, or C₂₋₆ alkynyl; or R₁ and R₃ on a sugar moiety can be linked to form —CH₂O, provided R₂ is hydrogen; R₄ is selected from the group consisting of —CH₂OH,

R₅ is selected from the group consisting of H, C₁₋₆ alkyl, —(CH₂CH₂O)_(p)CH₂CH₃, and —CH₂CH(OH)CH₂(O(CH₂)₂)_(n)CH₂NHC(O)R_(z); each R₆ is independently selected from H and NH₂; R_(b) is selected from C₁₋₆ alkyl, —OC₁₋₆ alkyl, halo, OH, CN, NO, NH₂, NHC₁₋₆alkyl, N(C₁₋₆alkyl)₂, SO₃H, SO₄, PO₄, C₁₋₃ haloalkyl, C₁₋₃ haloalkoxy, carboxyl, oxo, and S(O)_(g)alkyl; R is H or —P(O)(OH)_(2′); Q is independently selected from the group consisting of O, CH₂, and NH; R_(y) is —CH₂(O(CH₂)₂)_(n)(CH₂)_(k)NHC(O)—R_(z), or —(CH₂OCH₂)_(n)CH₂C(O)N((CH₂)_(k)NHC(O)R_(z))₂; R_(z) is cholesterol; g is 1, 2, or 3; k and n are independently selected from 1, 2, 3, 4, 5, and 6; p is an integer from 1 to 50. For purposes of this invention, the compound of Formula Ia and I comprises 24 nucleotides selected from the group consisting of 1, 2, 3, 4, 5, and 6,

wherein R₁, R₂, R₃, R₄, R₆, R_(x), R_(s), X, V, Y, K, Z, and W are as described herein. Adenine is present at positions 5, 9, 12, 17, 18, 19 and 22. Uracil is present at positions 3, 6, 7, 8, 16 and 20. When the hairpin is present, and reading from left to right, up to 10 self complementary base pairs are possible.

The compounds of the invention also include adenine derivative, inosine, or derivatives thereof, as depicted below:

An embodiment of the invention of Formula Ia and I is realized when it is read from left to right beginning with guanine (G), which is at the 5′ end, and ending with cytosine (C) at the 3′ end.

Another embodiment of the invention is compounds of formula (I), or a pharmaceutically acceptable salt thereof, wherein any X is independently selected from O and S, any Y is independently selected from OH and SH, any V is independently selected from O or S; and any K is independently selected from H and OH

In another embodiment, any R₁ is independently selected from the group consisting of H, halogen, —OH, —OMe, Me, —OCF₃, O(CH₂)₂OCH₃, NH₂, —OCH₂C≡CH, any R₂ is independently selected from the group consisting of H, halogen, —OH, —OMe, Me, and —OCF₃.

In another embodiment, any X is independently selected from O and S, any Y is independently selected from OH and SH, any V is independently selected from O or S; K is H, any W and Z within a single nine membered bicycle is independently selected from W═N and Z═—CH—; W═—CH— and Z═N and W═—CH— and Z is CH, any R₁ is selected from the group consisting of H, halogen, —OH, —OMe, Me, —OCF₃, —O(CH₂)₂OCH₃, —NH₂, —OCH₂C≡CH, any R₂ is independently selected from the group consisting of H, halogen, —OH, —OMe, -Me, and —OCF₃, and any R₃ is independently selected from the group consisting of H, halogen, C₁₋₆ alkyl, and —C≡CH.

An embodiment of the invention of Formula Ia and I is realized when any X is independently selected from O, S, and —CH₂—. A subembodiment of this aspect of the invention is realized when any X is independently selected from O and S. A subembodiment of this aspect of the invention is realized when each X is O. A subembodiment of this aspect of the invention is realized when each X is S. A subembodiment of this aspect of the invention is realized when each X is —CH₂—.

Another embodiment of the invention of Formula Ia and I is realized when any Y is independently selected from OH and SH A subembodiment of this aspect of the invention is realized when each Y is SH. A subembodiment of this aspect of the invention is realized when each Y is OH.

An embodiment of this aspect of the invention is realized when one to three Y is

A further subembodiment of this aspect of the invention is realized when the heteroaryl is optionally substituted triazolyl, said triazolyl optionally substituted with 1 to 3 groups of R_(b). Another embodiment of this aspect of the invention is realized when R_(y) is —CH₂(O(CH₂)₂)_(n)(CH₂)_(k)NHC(O)—R_(z). Still another subembodiment of this aspect of the invention is realized when R_(y) is —(CH₂OCH₂)_(n)CH₂C(O)N((CH₂)_(k)NHC(O)R_(z))₂. Another subembodiment of this aspect of the invention is realized when Y is

selected from

Another embodiment of the invention of Formula Ia and I is realized when V is O. Another subembodiment of the invention of Formula Ia and Formula I is realized when each V is S.

Still another embodiment of the invention of Formula Ia and Formula Ia and I is realized when K is H.

Another embodiment of the invention of Formula Ia and I is realized when K is

A subembodiment of this aspect of the invention is realized when R₅ is selected from the group consisting of H, C₁₋₆ alkyl and —(CH₂CH₂O)_(p)CH₂CH₃. Another subembodiment of this aspect of the invention is realized when R₅ is H. Another subembodiment of this aspect of the invention is realized when R₅ is C₁₋₆ alkyl. Still another subembodiment of this aspect of the invention is realized when R₅ is —(CH₂CH₂O)_(p)CH₂CH₃.

An embodiment of the invention of Formula Ia and I is realized when any W is independently selected from N and —CH—. Another embodiment of the invention of Formula Ia and I is realized when at least one W is N. Another embodiment of the invention of Formula Ia and I is realized when one to three W is N and the others are —CH—. Another embodiment of the invention of Formula Ia and I is realized when two or more W is —CH—. Another embodiment of the invention of Formula Ia and I is realized when one to three W is —CF—.

An embodiment of the invention of Formula Ia and I is realized when at least one Z is N. Still another embodiment of the invention of Formula Ia and I is realized when each Z is N. Another embodiment of the invention of Formula Ia and I is realized when each Z is —CH—. Another embodiment of the invention of Formula Ia and I is realized when at least one Z is —CH—. Another embodiment of the invention of Formula I is realized when any W and Z within a single nine membered bicycle is independently selected from W═N and Z═—CH—; W═—CH— and Z═N; W═N and Z═N; and W═—CH— and Z is —CH—. Another embodiment of the invention of Formula Ia and I is realized when each W is N and each Z is —CH—. Another embodiment of the invention of the Formula Ia and I is realized when each W is —CH— and each Z is —CH—. Another embodiment of the invention of the Formula Ia and I is realized when each W═—CH— and each Z═N. Another embodiment of the invention of the Formula Ia and I is realized when each W═N and each Z═N.

An embodiment of the invention of Formula Ia and I is realized when any R₁ of Formula Ia and I is selected from the group consisting of H, halogen, —OH, —OMe, Me, —OCF₃, —O(CH₂)₂OCH₃, —OCH₂C≡CH, —NH₂,

Another embodiment of the invention of Formula Ia and I is realized when any R₁ is independently selected from the group consisting of H, halogen, —OH, —OMe, -Me, —OCF₃, —O(CH₂)₂OCH₃, —OCH₂C≡CH, and —NH₂. Another embodiment of the invention of Formula Ia and I is realized when any R₁ is independently selected from H, halogen, —OH, —OMe, -Me, —OCF₃, —O(CH₂)₂OCH₃, and —NH₂.

An embodiment of the invention of Formula Ia and I is realized when any R₂ is independently selected from the group consisting of H, halogen, —OH, —OMe, -Me, and —OCF₃. A subembodiment of this invention of Formula Ia and I is realized when any R₂ is independently selected from the group consisting of H and -Me.

An embodiment of the invention of Formula Ia and I is realized when one to three adjacent R₁ and R₂ may combine to form ═CH₂.

Another embodiment of the invention of Formula Ia and I is realized when any R₃ is independently H, halogen, C₁₋₆ alkyl, —C≡CH. A sub-embodiment of the invention Formula 1 is realized when any R₃ is independently selected from H, F, Cl, methyl, ethyl, propyl, butyl hexyl and —C≡CH. A subembodiment of the invention of Formula Ia and I is realized when one to three R₁ and R₃ combine to form —CH₂O— and R₂ is hydrogen. A subembodiment of the invention of Formula I is realized when R₃ is hydrogen.

An embodiment of the invention of Formula Ia and I is realized when R₄ is —CH₂OH. Another embodiment of the invention of Formula Ia and I is realized when R₄ is

A subembodiment of this aspect of the invention is realized when R is H. Another subembodiment of the invention of Formula Ia and I is realized when R is —P(O)(OH)₂. A subembodiment of this aspect of the invention is realized when Q is O. Another subembodiment of this aspect of the invention is realized when Q is —NH—. Another subembodiment of this aspect of the invention is realized when Q is —CH₂—. Another subembodiment of this aspect of the invention is realized when g is 1, R is H and Q is O. Another subembodiment of the invention is realized when g is 2, R is H and Q is O. Still another subembodiment of the invention is realized when g is 3, R is H and Q is O. Yet another subembodiment of this aspect of the invention is realized when g is 1 and Q is —CH₂—, or —NH—. Another subembodiment of the invention is realized when g is 2, and Q is —NH— or —CH₂—. Still another subembodiment of the invention is realized when g is 3, and Q is —NH—, or —CH₂—.

Another embodiment of the invention of Formula Ia and I is realized when R₄ is

Another subembodiment of this aspect of the invention is realized when R is H. Another subembodiment of the invention of Formula Ia and I is realized when R is —P(O)(OH)₂. A subembodiment of this aspect of the invention is realized when Q is O. Another subembodiment of this aspect of the invention is realized when Q is NH. Another subembodiment of this aspect of the invention is realized when Q is CH₂. Another subembodiment of this aspect of the invention is realized when g is 1, R is H and Q is O. Another subembodiment of the invention is realized when g is 2, R is H and Q is O. Still another subembodiment of the invention is realized when g is 3, R is H and Q is O. Yet another subembodiment of this aspect of the invention is realized when g is 1 and Q is —CH₂—, or —NH—. Another subembodiment of the invention is realized when g is 2, and Q is —NH— or —CH₂—. Still another subembodiment of the invention is realized when g is 3, and Q is —NH—, or —CH₂—. Still another embodiment of the invention of Formula Ia and I is realized when R₄ is

An embodiment of the invention of Formula Ia and I is realized when R₄ is

An embodiment of the invention of Formula Ia and I is realized when R₅ is H. Another embodiment of the invention of Formula Ia and I is realized when R₅ is C₁₋₆ alkyl, wherein alkyl is selected from the group consisting of methyl, ethyl, propyl, butyl, pentyl and hexyl. Another embodiment of the invention of Formula Ia and I is realized when R₅ is —(CH₂CH₂O)_(p)CH₂CH₃. Yet another embodiment of the invention of Formula Ia and I is realized when R₅ is

An embodiment of the invention of Formula Ia and I is realized when R is H. Another embodiment of the invention of Formula Ia and I is realized when R is —P(O)(OH)₂.

An embodiment of the invention of Formula Ia and I is realized when R₄ is —CH₂OH and K is H. A subembodiment of this aspect of the invention is realized when Q is O.

Another embodiment of the invention of Formula Ia and I is realized when R₄ is —CH₂OH and K is P(O)(OH)₂. A subembodiment of this aspect of the invention is realized when Q is O.

An embodiment of the invention of Formula Ia and I is realized when one or more adjacent R₁ and R₂ combine to form ═CH₂, and R₄ is —CH₂OH. A subembodiment of this aspect of the invention is realized when K is H. Another subembodiment of this aspect of the invention is realized when K is —P(O)(OH)₂.

An embodiment of the invention of Formula Ia and I is realized when one or more adjacent R₁ and R₂ combine to form ═CH₂; and R₄ is selected from the group consisting of

Still another subembodiment of this aspect of the invention is realized when Q is O. Yet another subembodiment of this aspect of the invention is realized when Q is —NH—, —CH₂—. A subembodiment of this aspect of the invention is realized when K is H. Another subembodiment of this aspect of the invention is realized when K is —P(O)(OH)₂. Another subembodiment of this aspect of the invention is realized when R is H. Another subembodiment of the invention of Formula Ia and I is realized when R is —P(O)(OH)₂.

An embodiment of the invention of Formula Ia and I is realized when R₁ and R₃ on a sugar moiety are linked to form —CH₂O— and R₄ is —CH₂OH. A subembodiment of this aspect of the invention is realized when K is H. Another subembodiment of this aspect of the invention is realized when K is —P(O)(OH)₂.

An embodiment of the invention of Formula Ia and I is realized when R₁ and R₃ on a sugar moiety are linked to form —CH₂O and R₄ is selected from the group consisting of

A subembodiment of this aspect of the invention is realized when K is H. Another subembodiment of this aspect of the invention is realized when K is —P(O)(OH)₂. Still another subembodiment of this aspect of the invention is realized when Q is O. Yet another subembodiment of this aspect of the invention is realized when Q is independently selected from O, —CH₂— or —NH—, —CH₂—. Another subembodiment of this aspect of the invention is realized when R is H. Another subembodiment of the invention of Formula Ia and I is realized when R is —P(O)(OH)₂.

Another aspect of the invention of Formula Ia is realized when R_(x) is hydrogen. Still another aspect of the invention of Formula Ia is realized when R_(x) is —C≡CH.

Another aspect of the invention of Formula Ia is realized when R_(s) is NH₂. Still another aspect of the invention of Formula Ia is realized when R_(s) is —C(O)—.

As described herein, unless otherwise indicated, the use of a compound in treatment means that an amount of the compound, generally presented as a component of a formulation that comprises other excipients, is administered in aliquots of an amount, and at time intervals, which provide and maintain at least a therapeutic serum level of at least one pharmaceutically active form of the compound over the time interval between dose administrations.

When any variable (e.g. aryl, heterocycle, R¹, R² etc.) occurs more than one time in any constituent, its definition on each occurrence is independent of every other occurrence. Also, combinations of substituents/or variables are permissible only if such combinations result in stable compounds, is chemically feasible and/or valency permits.

As used herein, unless otherwise specified, the terms in the paragraphs immediately below have the indicated meaning.

“Alkyl” refers to both branched- and straight-chain saturated aliphatic hydrocarbon groups of 1 to 18 carbon atoms, or more specifically, 1 to 12 carbon atoms. Examples of such groups include, but are not limited to, methyl (Me), ethyl (Et), n-propyl (Pr), n-butyl (Bu), n-pentyl, n-hexyl, and the isomers thereof such as isopropyl (i-Pr), isobutyl (i-Bu), sec-butyl (s-Bu), tert-butyl (t-Bu), isopentyl, and isohexyl. Alkyl groups may be optionally substituted with one or more substituents as defined herein. “C₁₋₆alkyl” refers to an alkyl group as defined herein having 1 to 6 carbon atoms.

“Carboxyl” refers to a monovalent functional group or radical (e.g., COOH) typical of organic acids.

“Halo” or “halogen” refers to fluoro, chloro, bromo or iodo, unless otherwise noted.

The term “haloalkyl” refers to an alkyl in which one or more hydrogen atoms are replaced by halo, and includes alkyl moieties in which all hydrogens have been replaced by halo (e.g., perfluoroalkyl). Alkyl and haloalkyl groups may be optionally inserted with O, N, or S. The terms “aralkyl” refers to an alkyl moiety in which an alkyl hydrogen atom is replaced by an aryl group. Aralkyl includes groups in which more than one hydrogen atom has been replaced by an aryl group. Examples of “aralkyl” include benzyl, 9-fluorenyl, benzhydryl, and trityl groups.

The term “alkenyl” refers to a straight or branched hydrocarbon chain containing 2-8 carbon atoms and characterized in having one or more double bonds. Examples of a typical alkenyl include, but not limited to, allyl, propenyl, 2-butenyl, 3-hexenyl and 3-octenyl groups.

The term “alkynyl” refers to a straight or branched hydrocarbon chain containing 2-8 carbon atoms and characterized in having one or more triple bonds. Some examples of a typical alkynyl are ethynyl, 2-propynyl, and 3-methylbutynyl, and propargyl.

The term “alkoxy” refers to an alky group singularly bonded to oxygen, and the terms “cycloalkoxy” and “arylkoxy” refer to an cyclo alky and aryl alkyl groups singularly bonded to oxygen respectively.

The term “alkylene” refers to a divalent alkyl, e.g., —CH₂—, —CH₂CH₂—, and —CH₂CH₂CH₂—.

The term “cycloalkyl” as employed herein includes saturated cyclic, bicyclic, tricyclic, or polycyclic hydrocarbon groups having 3 to 12 carbons. The cycloalkyl groups herein described may also contain fused rings. Fused rings are rings that share a common carbon-carbon bond or a common carbon atom (e.g., spiro-fused rings). Examples of cycloalkyl moieties include, but are not limited to, cycylopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantyl, norbornyl, and decalin.

The term “heterocyclyl” refers to a nonaromatic 3-10 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ring system having 1-3 heteroatoms of monocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms selected from O, N, or S (e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S if monocyclic, bicyclic, or tricyclic, respectively). The heterocyclyl groups herein described may also contain fused rings. Fused rings are rings that share a common carbon-carbon bond or a common carbon atom (e.g., spiro-fused rings). Examples of heterocyclyl include, but are not limited to tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, morpholino, pyrrolinyl and pyrrolidinyl.

The term “cycloalkenyl” as employed herein includes partially unsaturated, nonaromatic, cyclic, bicyclic, tricyclic, or polycyclic hydrocarbon groups having 5 to 12 carbons, preferably 5 to 8 carbons. The cycloalkenyl groups herein described may also contain fused rings. Fused rings are rings that share a common carbon-carbon bond or a common carbon atom (e.g., spiro-fused rings). Examples of cycloalkenyl moieties include, but are not limited to, cyclohexenyl, and cyclohexadienyl.

The term “heteroaryl” refers to an aromatic 5-8 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ring system having 1-3 heteroatoms for monocyclic, 1-6heteroatoms for bicyclic, or 1-9 heteroatoms for tricyclic, said heteroatoms selected from O, N, or S (e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S for monocyclic, bicyclic, or tricyclic, respectively). The heteroaryl groups herein described may also contain fused rings that share a common carbon-carbon bond.

The term “oxo” refers to an oxygen atom, which forms a carbonyl when attached to carbon, an N-oxide when attached to nitrogen, and a sulfoxide or sulfone when attached to sulfur.

The term “acyl” refers to an alkylcarbonyl, cycloalkylcarbonyl, arylcarbonyl, heterocyclylcarbonyl, or heteroarylcarbonyl substituent.

The term “nucleobase” refers to a nitrogen-containing heterocyclic moiety, which are the parts of the nucleic acids that are involved in the hydrogen-bonding and bind one nucleic acid strand to another complementary strand in a sequence specific manner. The most common naturally-occurring nucleobases are: adenine (A), cyctosine (C), guanine (G), thymine (T), and uracil (U).

The term “modified nucleobase” refers to a moiety that can replace a nucleobase. The modified nucleobase mimics the spatial arrangement, electronic properties, or some other physiochemical property of the nucleobase and retains the property of the hydrogen-bonding that binds one nucleic acid strand to another in a sequence specific manner. A modified nucleobase can pair with at least one of the five naturally occurring bases (uracil, thymine, adenine, cytosine, and guanine) without substantially affecting the melting behavior, recognition by intracellular enzymes, or activity of the oligonucleotide duplex. The term “modified nucleoside” or “modified nucleotide” refers to a nucleoside or nucleotide that contains a modified nucleobase and/or other chemical modification disclosed herein, such as modified sugar, modified phosphorus atom bridges or modified internucleoside linkages. Non-limiting examples of nucleobases include uracil, thymine, adenine, cytosine, and guanine optionally having their respective amino groups protected by, e.g., acyl protecting groups, 5-propynyl-uracil, 2-thio-5-propynyl-uracil, 5-methylcytosine, 2-fluorouracil, 2-fluorocytosine, 5-bromouracil, 5-iodouracil, 2,6-diaminopurine, azacytosine, 2-thiouracil, 2-thiothymine, 2-aminopurine, N9-(2-amino-6-chloropurine, N9-(2,6-diaminopurine), hypoxanthine, N9-(7-deaza-guanine), N9-(7-deaza-8-aza-guanine), N8-(8-aza-7-deazaadenine), pyrimidine analogs such as pseudoisocytosine and pseudouracil and other modified nucleobases such as 8-substituted purines, xanthine, or hypoxanthine.

The terms “adeninyl, cytosinyl, guaninyl, thyminyl, and uracilyl” and the like refer to radicals of adenine, cytosine, guanine, thymine, and uracil.

The term cholesterol is presented by

and when used as a radical, it is linked to the core structure through bond having the “OH” group. A “protected” moiety refers to a reactive functional group, e.g., a hydroxyl group or an amino group, or a class of molecules, e.g., sugars, having one or more functional groups, in which the reactivity of the functional group is temporarily blocked by the presence of an attached protecting group. Protecting groups useful for the monomers and methods described herein can be found, e.g., in Greene, T. W., Protective Groups in Organic Synthesis (John Wiley and Sons: New York), 1981, which is hereby incorporated by reference.

A further aspect of the present invention relates to a pharmaceutical composition comprising a modified oligonucleotide as defined herein.

The pharmaceutical composition according to the invention may further comprise pharmaceutically acceptable carriers, diluents, and/or adjuvants. The term “carrier” when used herein includes carriers, excipients and/or stabilisers that are non-toxic to the cell or mammal being exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carriers are aqueous pH buffered solutions or liposomes. Examples of physiologically acceptable carriers include buffers such as phosphate, citrate and other organic acids (however, with regard to the formulation of the present invention, a phosphate buffer is preferred); anti-oxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinyl pyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose or dextrins, gelating agents such as EDTA, sugar, alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and/or non-ionic surfactants such as TWEEN, polyethylene or polyethylene glycol. According to an embodiment the compound of the invention is dissolved in sterile deionized water.

Such a composition and/or formulation according to the invention can be administered to a subject in need thereof, particularly a human patient, in a sufficient dose for the treatment of the specific conditions by suitable means. For example, the composition and/or formulation according to the invention may be formulated as a pharmaceutical composition together with pharmaceutically acceptable carriers, diluents and/or adjuvants. Therapeutic efficiency and toxicity may be determined according to standard protocols. The pharmaceutical composition may be administered intratumorally, and systemically, e.g., intraperitoneally, intramuscularly, intravenously, locally, such as intranasally, subcutaneously, intradermally or intrathecally. The dose of the composition and/or formulation administered will, of course, be dependent on the subject to be treated and on the condition of the subject, such as the subject's weight, the subject's age and the type and severity of the disease or injury to be treated, the manner of administration and the judgement of the prescribing physician.

In an embodiment, the pharmaceutical composition is administered intradermally. In another embodiment, the composition is administered intradermally via tattooing, microneedling and/or microneedle patches.

Methods of Use

Compounds described herein having therapeutic applications, such as the compounds of general Formula Ia and Formula I, and the compounds of Examples 1 through 838, can be administered to a patient for the purpose of inducing an immune response, inducing a RIG-I-dependent IFN production and/or inducing anti-tumor activity. In one embodiment, the present invention provides for both prophylactic and therapeutic methods of inducing an IFN response in a patient. The term “administration” and variants thereof (e.g., “administering” a compound) means providing the compound to the individual in need of treatment. When a compound is provided in combination with one or more other active agents (e.g., antiviral agents useful for treating HCV infection or anti-tumor agents for treating cancers), “administration” and its variants are each understood to include concurrent and sequential provision of the compound or salt and other agents.

The present invention encompasses the use of the short oligonucleotide hairpin molecule of the invention to prevent and/or treat any disease, disorder, or condition in which inducing IFN production would be beneficial. For example, increased IFN production, by way of the nucleic acid molecule of the invention, may be beneficial to prevent or treat a wide variety of disorders, including, but not limited to, bacterial infection, viral infection, parasitic infection, immune disorders, respiratory disorders, cancer and the like.

Infections include, but are not limited to, viral infections, bacterial infections, anthrax, parasitic infections, fungal infections and prion infection.

Viral infections include, but are not limited to, infections by hepatitis C, hepatitis B, influenza virus, herpes simplex virus (HSV), human immunodeficiency virus (HIV), respiratory syncytial virus (RSV), vesicular stomatitis virus (VSV), cytomegalovirus (CMV), poliovirus, encephalomyocarditis virus (EMCV), human papillomavirus (HPV) and smallpox virus. In one embodiment, the infection is an upper respiratory tract infection caused by viruses and/or bacteria, in particular, flu, more specifically, bird flu.

Bacterial infections include, but are not limited to, infections by streptococci, staphylococci, E. coli, and Pseudomonas. In one embodiment, the bacterial infection is an intracellular bacterial infection which is an infection by an intracellular bacterium such as mycobacteria (tuberculosis), chlamydia, mycoplasma, listeria, and an facultative intracellular bacterium such as Staphylococcus aureus.

Parasitic infections include, but are not limited to, worm infections, in particular, intestinal worm infection, microeukaryotes, and vector-borne diseases, including for example Leishmaniasis.

In a preferred embodiment, the infection is a viral infection or an intracellular bacterial infection. In a more preferred embodiment, the infection is a viral infection by hepatitis C, hepatitis B, influenza virus, RSV, HPV, HSV1, HSV2, and CMV.

Immune disorders include, but are not limited to, allergies, autoimmune disorders, and immunodeficiencies.

Allergies include, but are not limited to, respiratory allergies, contact allergies and food allergies.

Autoimmune diseases or disorders include, but are not limited to, multiple sclerosis, diabetes mellitus, arthritis (including rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, psoriatic arthritis), encephalomyelitis, myasthenia gravis, systemic lupus erythematosis, autoimmune thyroiditis, dermatisis (including atopic dermatitis and eczematous dermatitis), psoriasis, Siogren's Syndrome, Crohn's Disease, aphthous ulcer, iritis, conjunctivitis, keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma, cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, drug eruptions, leprosy reversal reactions, erythema nodosum leprosum, autoimmune uveitis, allergic encephalomyelitis, acute necrotizing hemorrhagic encephalopathy, idiopathic bilateral progressive sensorineural hearing loss, aplastic anemia, pure red cell anemia, idiopathic thrombocytopenia, polychondritis, Wegener's granulomatosis, chronic active hepatitis, Stevens-Johnson syndrome, idiopathic sprue, lichen planus, Graves' disease, sarcoidosis, primary biliary cirrhosis, uveitis posterior, and interstitial lung fibrosis.

Immunodeficiencies include, but are not limited to, spontaneous immunodeficiency, acquired immunodeficiency (including AIDS), drug-induced immunodeficiency or immunosuppression (such as that induced by immunosuppressants used in transplantation and chemotherapeutic agents used for treating cancer), and immunosuppression caused by chronic hemodialysis, trauma or surgical procedures.

Respiratory disorders include, but are not limited to, acute lung injury (ALI), acute respiratory distress syndrome (ARDS), asthma, chronic obstructive pulmonary disease (COPD), obstructive sleep apnea (OSA), idiopathic pulmonary fibrosis (IPF), tuberculosis, pulmonary hypertension, pleural effusion, and lung cancer.

Examples of cancers include, but are not limited to, Acute Lymphoblastic Leukemia; Acute Myeloid Leukemia; Adrenocortical Carcinoma; AIDS-Related Lymphoma; AIDS-Related Malignancies; Anal Cancer; Astrocytoma; Bile Duct Cancer; Bladder Cancer; Bone Cancer, Osteosarcoma/Malignant Fibrous Histiocytoma; Brain Stem Glioma; Brain Tumor, Cerebellar Astrocytoma; Brain Tumor, Cerebral Astrocytoma/Malignant Glioma; Brain Tumor, Ependymoma; Brain Tumor, Medulloblastoma; Brain Tumor, Supratentorial Primitive Neuroectodermal Tumors; Brain Tumor, Visual Pathway and Hypothalamic Glioma; Breast Cancer; Bronchial Adenomas/Carcinoids; Carcinoid Tumor; Carcinoid Tumor, Gastrointestinal; Carcinoma, Adrenocortical; Carcinoma, Islet Cell; Central Nervous System Lymphoma, Primary; Cerebral Astrocytoma/Malignant Glioma; Cervical Cancer; Chronic Lymphocytic Leukemia; Chronic Myelogenous Leukemia; Chronic Myeloproliferative Disorders; Clear Cell Sarcoma of Tendon Sheaths; Colon Cancer; Colorectal Cancer; Cutaneous T-Cell Lymphoma; Endometrial Cancer; Ependymoma; Epithelial Cancer, Ovarian; Esophageal Cancer; Esophageal Cancer; Ewing's Family of Tumors; Extracranial Germ Cell Tumor; Extrahepatic Bile Duct Cancer; Eye Cancer, Intraocular Melanoma; Eye Cancer, Retinoblastoma; Gallbladder Cancer; Gastric (Stomach) Cancer; Gastrointestinal Carcinoid Tumor; Germ Cell Tumor, Extracranial, Childhood; Germ Cell Tumor, Extragonadal; Germ Cell Tumor, Ovarian; Gestational Trophoblastic Tumor; Glioma, Childhood Brain Stem; Glioma, Childhood Visual Pathway and Hypothalamic; Hairy Cell Leukemia; Head and Neck Cancer; Hepatocellular (Liver) Cancer; Hodgkin's Lymphoma; Hypopharyngeal Cancer; Hypothalamic and Visual Pathway Glioma; Intraocular Melanoma; Islet Cell Carcinoma (Endocrine Pancreas); Kaposi's Sarcoma; Kidney Cancer; Laryngeal Cancer; Leukemia, Acute Lymphoblastic; Leukemia, Acute Myeloid; Leukemia, Chronic Lymphocytic; Leukemia, Chronic Myelogenous; Leukemia, Hairy Cell; Lip and Oral Cavity Cancer; Liver Cancer; Lung Cancer, Non-Small Cell; Lung Cancer, Small Cell; Lymphoblastic Leukemia; Lymphoma, AIDS-Related; Lymphoma, Central Nervous System (Primary); Lymphoma, Cutaneous T-Cell; Lymphoma, Hodgkin's; Lymphoma, Hodgkin's During Pregnancy; Lymphoma, Non-Hodgkin's; Lymphoma, Primary Central Nervous System; Macroglobulinemia, Waldenstrom's; Male Breast Cancer; Malignant Mesothelioma; Malignant Thymoma; Medulloblastoma, Childhood; Melanoma; Melanoma, Intraocular; Merkel Cell Carcinoma; Mesothelioma, Malignant; Metastatic Squamous Neck Cancer with Occult Primary; Multiple Endocrine Neoplasia Syndrome, Childhood; Multiple Myeloma/Plasma Cell Neoplasm; Mycosis Fungoides; Myelodysplastic Syndromes; Myelogenous Leukemia, Chronic; Myeloid Leukemia; Myeloma, Multiple; Myeloproliferative Disorders, Chronic; Nasal Cavity and Paranasal Sinus Cancer; Nasopharyngeal Cancer; Neuroblastoma; Non-Hodgkin's Lymphoma; Non-Small Cell Lung Cancer; Oral Cancer; Oral Cavity and Lip Cancer; Oropharyngeal Cancer; steosarcoma/Malignant Fibrous Histiocytoma of Bone; Ovarian Epithelial Cancer; Ovarian Germ Cell Tumor; Ovarian Low Malignant Potential Tumor; Pancreatic Cancer; Paranasal Sinus and Nasal Cavity Cancer; Parathyroid Cancer; Penile Cancer; Pheochromocytoma; Pineal and Supratentorial Primitive Neuroectodermal Tumors; Pituitary Tumor; Plasma Cell Neoplasm/Multiple Myeloma; Pleuropulmonary Blastoma; Pregnancy and Breast Cancer; Pregnancy and Hodgkin's Lymphoma; Pregnancy and Non-Hodgkin's Lymphoma; Primary Central Nervous System Lymphoma; Primary Liver Cancer; Prostate Cancer; Rectal Cancer; Renal Cell (Kidney) Cancer; Renal Pelvis and Ureter, Transitional Cell Cancer; Retinoblastoma; Rhabdomyosarcoma; Salivary Gland Cancer; Sarcoma, Ewing's Family of Tumors; Sarcoma, Kaposi's; Sarcoma (Osteosarcoma)/Malignant Fibrous Histiocytoma of Bone; Sarcoma, Soft Tissue; Sezary Syndrome; Skin Cancer; Skin Cancer (Melanoma); Skin Carcinoma, Merkel Cell; Small Cell Lung Cancer; Small Intestine Cancer; Soft Tissue Sarcoma; Squamous Neck Cancer with Occult Primary, Metastatic; Stomach (Gastric) Cancer; Supratentorial Primitive Neuroectodermal Tumors; T-Cell Lymphoma, Cutaneous; Testicular Cancer; Thymoma, Malignant; Thyroid Cancer; Transitional Cell Cancer of the Renal Pelvis and Ureter; Trophoblastic Tumor, Gestational; Ureter and Renal Pelvis, Transitional Cell Cancer; Urethral Cancer; Uterine Sarcoma; Vaginal Cancer; Visual Pathway and Hypothalamic Glioma; Vulvar Cancer; Waldenstrom's Macro globulinemia; and Wilms' Tumor.

In one embodiment, the cancer is brain cancer, such as an astrocytic tumor (e.g., pilocytic astrocytoma, subependymal giant-cell astrocytoma, diffuse astrocytoma, pleomorphic xanthoastrocytoma, anaplastic astrocytoma, astrocytoma, giant cell glioblastoma, glioblastoma, secondary glioblastoma, primary adult glioblastoma, and primary pediatric glioblastoma); oligodendroglial tumor (e.g., oligodendroglioma, and anaplastic oligodendroglioma); oligoastrocytic tumor (e.g., oligoastrocytoma, and anaplastic oligoastrocytoma); ependymoma (e.g., myxopapillary ependymoma, and anaplastic ependymoma); medulloblastoma; primitive neuroectodermal tumor, schwannoma, meningioma, meatypical meningioma, anaplastic meningioma; and pituitary adenoma. In another embodiment, the brain cancer is glioma, glioblastoma multiforme, paraganglioma, or suprantentorial primordial neuroectodermal tumors (sPNET).

In another embodiment, the cancer is leukemia, such as acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), chronic myelogenous leukemia (CML), myeloproliferative neoplasm (MPN), post-MPN AML, post-MDS AML, del(5q)-associated high risk MDS or AML, blast-phase chronic myelogenous leukemia, angioimmunoblastic lymphoma, and acute lymphoblastic leukemia.

In one embodiment, the cancer is skin cancer, including melanoma. In another embodiment, the cancer is prostate cancer, breast cancer, thyroid cancer, colon cancer, or lung cancer. In another embodiment, the cancer is sarcoma, including central chondrosarcoma, central and periosteal chondroma, and fibrosarcoma. In another embodiment, the cancer is cholangiocarcinoma.

Administration and Dosing

As used herein, the terms “treatment” and “treating” refer to all processes wherein there may be a slowing, interrupting, arresting, controlling, or stopping of the progression of a disease or disorder described herein. The terms do not necessarily indicate a total elimination of all disease or disorder symptoms. The terms also include the potential prophylactic therapy of the mentioned conditions, particularly in a subject that is predisposed to such disease or disorder.

The terms “administration of” and or “administering” a compound should be understood to include providing a compound described herein, or a pharmaceutically acceptable salt thereof, and compositions of the foregoing to a subject.

The amount of a compound administered to a subject is an amount sufficient to induce an immune response and/or to induce RIG-I-dependent type I interferon production in the subject. In an embodiment, the amount of a compound can be an “effective amount” or “therapeutically effective amount,” wherein the subject compound is administered in an amount that will elicit a biological or medical response of a tissue, system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician. An effective amount does not necessarily include considerations of toxicity and safety related to the administration of a compound.

An effective amount of a compound will vary with the particular compound chosen (e.g., considering the potency, efficacy, and/or half-life of the compound); the route of administration chosen; the condition being treated; the severity of the condition being treated; the age, size, weight, and physical condition of the subject being treated; the medical history of the subject being treated; the duration of the treatment; the nature of a concurrent therapy; the desired therapeutic effect; and like factors and can be routinely determined by the skilled artisan.

Biological methods for introducing a nucleic acid into a host cell include the use of DNA and RNA vectors. Viral vectors, and especially retroviral vectors, have become the most widely used method for inserting genes into mamma-lian, e.g., human cells.

The compounds disclosed herein may be administered by any suitable route including oral and parenteral administration. Parenteral administration is typically by injection or infusion and includes intravenous, intramuscular, and subcutaneous injection or infusion.

The compounds disclosed herein may be administered once or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. For example, doses may be administered one, two, three, or four times per day. Doses may be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. Suitable dosing regimens for a compound disclosed herein depend on the pharmacokinetic properties of that compound, such as absorption, distribution and half-life which can be determined by a skilled artisan. In addition, suitable dosing regimens, including the duration such regimens are administered, for a compound disclosed herein depend on the disease or condition being treated, the severity of the disease or condition, the age and physical condition of the subject being treated, the medical history of the subject being treated, the nature of concurrent therapy, the desired therapeutic effect, and like factors within the knowledge and expertise of the skilled artisan. It will be further understood by such skilled artisans that suitable dosing regimens may require adjustment given an individual subject's response to the dosing regimen or over time as the individual subject needs change. Typical daily dosages may vary depending upon the particular route of administration chosen. Typical daily dosages for oral administration, to a human weighing approximately 70 kg would range from about 0.1 mg to about 2 g, or more specifically, 0.1 mg to 500 mg, or even more specifically, 0.2 mg to 100 mg, of a compound of general Formula Ia and Formula I.

One embodiment of the present disclosure provides for a method of treating a cell proliferation disorder comprising administration of an effective amount of a compound of general Formula Ia and Formula I to a subject in need of treatment thereof. In one embodiment, the cell proliferation disorder is cancer.

In one embodiment, the cancer is brain cancer, leukemia, skin cancer, prostate cancer, thyroid cancer, colon cancer, lung cancer or sarcoma. In another embodiment the cancer is glioma, glioblastoma multiforme, paraganglioma, suprantentorial primordial neuroectodermal tumors, acute myeloid leukemia, myelodysplastic syndrome, chronic myelogenous leukemia, melanoma, breast, prostate, thyroid, colon, lung, central chondrosarcoma, central and periosteal chondroma tumors, fibrosarcoma, and cholangiocarcinoma.

In one embodiment, disclosed herein is the use of a compound of general Formula Ia and Formula I in a therapy. The compound may be useful in a method of inducing an immune response and/or inducing RIG-I-dependent type I interferon production in a subject, such as a mammal in need of such inhibition, comprising administering an effective amount of the compound to the subject.

In one embodiment, disclosed herein is a pharmaceutical composition comprising at least one compound of general Formula Ia and Formula I, or a pharmaceutically acceptable salt thereof, for use in potential treatment to induce an immune response and/or to induce RIG-I-dependent type I interferon production.

In one embodiment, disclosed herein is the use of a compound of general Formula Ia and Formula I in the manufacture of a medicament for the treatment to induce an immune response and/or to induce RIG-I-dependent type I interferon production. In one embodiment, the disease or disorder to be treated is a cell proliferation disorder. In another embodiment, the cell proliferation disorder is cancer. In another embodiment, the cancer is brain cancer, leukemia, skin cancer, breast, prostate cancer, thyroid cancer, colon cancer, lung cancer, or sarcoma. In another embodiment, the cancer is glioma, glioblastoma multiforme, paraganglioma, suprantentorial primordial neuroectodermal tumors, acute myeloid leukemia, myelodysplastic syndrome, chronic myelogenous leukemia, melanoma, breast, prostate, thyroid, colon, lung, central chondrosarcoma, central and periosteal chondroma tumors, fibrosarcoma, and/or cholangiocarcinoma.

Compositions

The present invention provides a pharmaceutical composition comprising at least one nucleic acid molecule of the present invention and a pharmaceutically acceptable carrier. The formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient into association with a carrier or one or more other accessory ingredients, and then, if necessary or desirable, shaping or packaging the product into a desired single- or multi-dose unit.

The term “composition” as used herein is intended to encompass a dosage form comprising a specified compound in a specified amount, as well as any dosage form which results, directly or indirectly, from combination of a specified compound in a specified amount. Such term is intended to encompass a dosage form comprising a compound of general Formula Ia and Formula I, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients. Accordingly, the compositions of the present disclosure encompass any composition made by admixing a compound of the present disclosure and one or more pharmaceutically acceptable carrier or excipients. By “pharmaceutically acceptable”, it is meant the carriers or excipients are compatible with the compound disclosed herein and with other ingredients of the composition.

For the purpose of inducing an immune response and/or inducing a RIG-I-dependent type I interferon production, the compounds, optionally in the form of a salt, can be administered by means that produce contact of the active agent with the agent's site of action. They can be administered by conventional means available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or in a combination of therapeutic agents. They can be administered alone, but typically are administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice.

An embodiment of the invention is realized by a composition comprising a nucleic acid capable of inducing interferon production, wherein the molecule comprises a single strand of 24 bases by 5′ GCUCAUUUACCAGCGUAAAUGAGC 3′ (SEQ ID NO: 841), said single strand having a double-stranded section of less than 12 base pairs. A subembodiment of this aspect of the invention is realized when the single stranded nucleic acid forms a hairpin structure comprising the double-stranded section and a loop. Another subembodiment of this aspect of the invention is realized when the double-stranded nucleic acid comprises one or two blunt ends. Another subembodiment of this aspect of the invention is realized when

the double-stranded nucleic acid comprises a 5′ blunt end. Another subembodiment of this aspect of the invention is realized when the nucleic acid comprises at least one of the group consisting of a 5′-OH, 5′-triphosphate and a 5′-diphosphate. Another subembodiment of this aspect of the invention is realized when the molecule comprises a modified phosphodiester backbone. Still another subembodiment of this aspect of the invention is realized when the molecule comprises at least one 2′-modified nucleotide. A further aspect of this embodiment of the invention is realized when the 2′-modified nucleotide comprises a modification selected from the group consisting of: 2′-OH, 2′-methyl, 2′-OCF₃, 2′-OCH₂C≡CH, —NH₂, 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyl-oxyethyl (2′-O-DMAEOE), and 2′-O—N-methylacetamido (2′-O-NMA). Another subembodiment of this aspect of the invention is realized when the molecule comprises at least one modified phosphate group. Another subembodiment of this aspect of the invention is realized when the molecule comprises at least one modified base. Another subembodiment of this aspect of the invention is realized when the double stranded section comprises one or more mispaired bases.

The compositions of the disclosure are prepared using techniques and methods known to those skilled in the art. Some methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

In one embodiment, disclosed herein is a composition comprising a compound of general Formula Ia and Formula I or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients. The composition may be prepared and packaged in bulk form wherein an effective amount of a compound of the disclosure can be extracted and then given to a subject, such as with powders or syrups. Alternatively, the composition may be prepared and packaged in unit dosage form wherein each physically discrete unit contains an effective amount of a compound of general Formula Ia and Formula I. When prepared in unit dosage form, the composition of the disclosure typically contains from about 0.1 mg to 2 g, or more specifically, 0.1 mg to 500 mg, or even more specifically, 0.2 mg to 100 mg, of a compound of general Formula Ia and Formula I, or a pharmaceutically acceptable salt thereof.

The pharmaceutical composition according to the invention may further comprise pharmaceutically acceptable carriers, diluents, and/or adjuvants. The term “carrier” when used herein includes excipients and/or stabilisers that are non-toxic to the cell or mammal being exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carriers are aqueous pH buffered solutions or liposomes. Suitable pharmaceutically acceptable carriers or excipients include the following non-limiting types: diluents, lubricants, binders, disintegrants, fillers, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavoring agents, flavor masking agents, coloring agents, anti-caking agents, hemectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents. Particular examples of physiologically acceptable carriers include buffers such as phosphate, citrate and other organic acids; anti-oxidants including ascorbic acid, low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatine or immunoglobulins; hydrophilic polymers such as polyvinyl pyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose or dextrins; gelating agents such as EDTA, sugar, alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and/or non-ionic surfactants such as TWEEN, polyethylene or polyethylene glycol. In another embodiment the compound of the invention is dissolved in sterile deionized water.

A skilled artisan possesses the knowledge and skill in the art to select suitable pharmaceutically acceptable carriers and excipients in appropriate amounts for the use in the disclosure. In addition, there are a number of resources available to the skilled artisan, which describe pharmaceutically acceptable carriers and excipients and may be useful in selecting suitable pharmaceutically acceptable carriers and excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

The compounds disclosed herein and a pharmaceutically acceptable carrier or excipient(s) will typically be formulated into a dosage form adapted for administration to a subject by a desired route of administration. Many systems for administering active substances into cells are already known and include lipid nanoparticles (LNPs), emulsions, liposomes, exosomes, dendrimers, natural polymers, e.g., chitosan, polyethylenimines (PEIs), immuno- and ligand complexes, micelles, microparticles and/or polymeric nanoparticles and cyclodextrins (see, Drug Transport in Antimicrobial and Anticancer Chemotherapy. G. Papadakou Ed., CRC Press, 1995 and US2013/0037977). These systems may be adapted for (1) oral administration, such as tablets, capsules, caplets, pills, troches, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets, and cachets; and (2) parenteral administration, such as sterile solutions, suspensions, and powders for reconstitution. Suitable pharmaceutically acceptable carriers or excipients will vary depending upon the particular dosage form chosen. In addition, suitable pharmaceutically acceptable carriers or excipients may be chosen for a particular function that they may serve in the composition. For example, certain pharmaceutically acceptable carriers or excipients may be chosen for their ability to facilitate the production of uniform dosage forms. Certain pharmaceutically acceptable carriers or excipients may be chosen for their ability to facilitate the production of stable dosage forms. Certain pharmaceutically acceptable carriers or excipients may be chosen for their ability to facilitate the carrying or transporting of a compound disclosed herein, once administered to the subject, from one organ or portion of the body to another organ or another portion of the body. Certain pharmaceutically acceptable carriers or excipients may be chosen for their ability to enhance patient compliance.

In an embodiment, the disclosure is directed to a solid oral dosage form such as a tablet or capsule comprising an effective amount of a compound of the disclosure and a diluent or filler. Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g., corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives, (e.g., microcrystalline cellulose), calcium sulfate, and dibasic calcium phosphate. The oral solid dosage form may further comprise a binder. Suitable binders include starch (e.g., corn starch, potato starch, and pre-gelatinized starch) gelatin, acacia, sodium alginate, alginic acid, tragacanth, guar gum, povidone, and cellulose and its derivatives (e.g., microcrystalline cellulose). The oral solid dosage form may further comprise a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmelose, alginic acid, and sodium carboxymethyl cellulose. The oral solid dosage form may further comprise a lubricant. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate, and talc.

Where appropriate, dosage unit formulations for oral administration can be microencapsulated. The composition can also be prepared to prolong or sustain the release as, for example, by coating or embedding particulate material in polymers, wax, or the like.

The compounds disclosed herein may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyrancopolymer, polyhydroxypropylmethacrylamidephenol, polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysine substituted with palmitoyl residues. Furthermore, the compounds of the disclosure may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example polylactic acid, polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanacrylates and cross-linked or amphipathic block copolymers of hydrogels.

In one embodiment, the disclosure is directed to a liquid oral dosage form. Oral liquids such as solution, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of a compound disclosed herein. Syrups can be prepared by dissolving the compound of the disclosure in a suitably flavored aqueous solution; while elixirs are prepared through the use of a non-toxic alcoholic vehicle. Suspensions can be formulated by dispersing a compound disclosed herein in a non-toxic vehicle. Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives, flavor additives such as peppermint oil or other natural sweeteners or saccharin or other artificial sweeteners and the like can also be added.

In another embodiment, the disclosure is directed to compositions for parenteral administration. Compositions adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions that may include suspending agents and thickening agents. The compositions may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.

Combinations

The compounds of general Formula Ia and Formula I may be administered in combination with one or more additional therapeutic agents. In embodiments, one or more compound of general Formula Ia and Formula I may be co-adminstered. The additional therapeutic agent(s) may be administered in a single dosage form with the compound of general Formula Ia and Formula I, or the additional therapeutic agent(s) may be administered in separate dosage form(s) from the dosage form containing the compound of general Formula Ia and Formula I. The additional therapeutic agent(s) may be one or more agents selected from the group consisting of anti-viral compounds, antigens, adjuvants, anti-cancer agents, CTLA-4, LAG-3 and PD-1 pathway antagonists, lipids, peptides, chemotherapeutic agents, immunomodulatory cell lines, checkpoint inhibitors, vascular endothelial growth factor (VEGF) receptor inhibitors, topoisomerase II inhibitors, smoothen inhibitors, alkylating agents, anti-tumor antibiotics, anti-metabolites, retinoids, and immunomodulatory agents including but not limited to anti-cancer vaccines. It will be understood the descriptions of the above additional therapeutic agents may be overlapping. It will also be understood that the treatment combinations are subject to optimization, and it is understood that the best combination to use of the compounds of general Formula Ia and Formula I and one or more additional therapeutic agents will be determined based on the individual patient needs.

A compound disclosed herein may be used in combination with one or more other active agents, including but not limited to, other anti-cancer agents that are used in the prevention, treatment, control, amelioration, or reduction of risk of a particular disease or condition (e.g., cell proliferation disorders). In one embodiment, a compound disclosed herein is combined with one or more other anti-cancer agents for use in the prevention, treatment, control amelioration, or reduction of risk of a particular disease or condition for which the compounds disclosed herein are useful. Such other active agents may be administered, by a route and in an amount commonly used therefor, contemporaneously or sequentially with a compound of the present disclosure.

When a compound disclosed herein is used contemporaneously with one or more other active agents, a composition containing such other active agents in addition to the compound disclosed herein is contemplated. Accordingly, the compositions of the present disclosure include those that also contain one or more other active ingredients, in addition to a compound disclosed herein. A compound disclosed herein may be administered either simultaneously with, or before or after, one or more other therapeutic agent(s). A compound disclosed herein may be administered separately, by the same or different route of administration, or together in the same pharmaceutical composition as the other agent(s).

Products provided as a combined preparation include a composition comprising a compound of general Formula Ia and Formula I and one or more other active agent(s) together in the same pharmaceutical composition, or a compound of general Formula Ia and Formula I and one or more other therapeutic agent(s) in separate form, e.g. in the form of a kit.

The weight ratio of a compound disclosed herein to a second active agent may be varied and will depend upon the effective dose of each agent. Generally, an effective dose of each will be used. Thus, for example, when a compound disclosed herein is combined with another agent, the weight ratio of the compound disclosed herein to the other agent will generally range from about 1000:1 to about 1:1000, such as about 200:1 to about 1:200. Combinations of a compound disclosed herein and other active agents will generally also be within the aforementioned range, but in each case, an effective dose of each active agent should be used. In such combinations, the compound disclosed herein and other active agents may be administered separately or in conjunction. In addition, the administration of one element may be prior to, concurrent to, or subsequent to the administration of other agent(s).

In one embodiment, this disclosure provides a composition comprising a compound of general Formula Ia and Formula I and at least one other therapeutic agent as a combined preparation for simultaneous, separate or sequential use in therapy. In one embodiment, the therapy is the treatment of a cell proliferation disorder, such as cancer.

In one embodiment, the disclosure provides a kit comprising two or more separate pharmaceutical compositions, at least one of which contains a compound of general Formula Ia and Formula I. In one embodiment, the kit comprises means for separately retaining said compositions, such as a container, divided bottle, or divided foil packet. An example of such a kit is a blister pack, as typically used for the packaging of tablets, capsules, and the like.

A kit of this disclosure may be used for administration of different dosage forms, for example, oral and parenteral, for administration of the separate compositions at different dosage intervals, or for titration of the separate compositions against one another. To assist with compliance, a kit of the disclosure typically comprises directions for administration.

Disclosed herein is a use of a compound of general Formula Ia and Formula I for treating a cell proliferation disorder, wherein the medicament is prepared for administration with another active agent. The disclosure also provides the use of another active agent for treating a cell proliferation disorder, wherein the medicament is administered with a compound of general Formula Ia and Formula I.

The disclosure also provides the use of a compound of general Formula Ia and Formula I for treating a cell proliferation disorder, wherein the patient has previously (e.g., within 24 hours) been treated with another active agent. The disclosure also provides the use of another therapeutic agent for treating a cell proliferation disorder, wherein the patient has previously (e.g., within 24 hours) been treated with a compound of general Formula Ia and Formula I. The second agent may be applied a week, several weeks, a month, or several months after the administration of a compound disclosed herein.

Anti-viral compounds that may be used in combination with the compounds of general Formula Ia and Formula I disclosed herein include hepatitis B virus (HBV) inhibitors, hepatitis C virus (HCV) protease inhibitors, HCV polymerase inhibitors, HCV NS4A inhibitors, HCV NS5A inhibitors, HCV NS5b inhibitors, and human immunodeficiency virus (HIV) inhibitors.

Antigens and adjuvants that may be used in combination with the compounds of general Formula Ia and Formula I disclosed herein include B7 costimulatory molecule, interleukin-2, interferon-γ, GM-CSF, CTLA-4 antagonists, OX-40/OX-40 ligand, CD40/CD40 ligand, sargramostim, levamisol, vaccinia virus, Bacille Calmette-Guerin (BCG), liposomes, alum, Freund's complete or incomplete adjuvant, detoxified endotoxins, mineral oils, surface active substances such as lipolecithin, pluronic polyols, polyanions, peptides, and oil or hydrocarbon emulsions. Adjuvants, such as aluminum hydroxide or aluminum phosphate, can be added to increase the ability of the compound to trigger, enhance, or prolong an immune response. Additional materials, such as cytokines, chemokines, and bacterial nucleic acid sequences, like CpG, a toll-like receptor (TLR) 9 agonist as well as additional agonists for TLR 2, TLR 4, TLR 5, TLR 7, TLR 8, TLR9, including lipoprotein, LPS, monophosphoryllipid A, lipoteichoic acid, imiquimod, resiquimod, stimulator of interferon genes (STING) agonists and in addition retinoic acid-inducible gene I (RIG-I) agonists such as poly I:C, used separately or in combination with the described compositions are also potential adjuvants.

CLTA-4 and PD-1 pathways are important negative regulators of immune response. Activated T-cells upregulate CTLA-4, which binds on antigen-presenting cells and inhibits T-cell stimulation, IL-2 gene expression, and T-cell proliferation; these anti-tumor effects have been observed in mouse models of colon carcinoma, metastatic prostate cancer, and metastatic melanoma. PD-1 binds to active T-cells and suppresses T-cell activation; PD-1 antagonists have demonstrated anti-tumor effects as well. CTLA-4 and PD-1 pathway antagonists that may be used in combination with the compounds of general Formula Ia and Formula I disclosed herein include ipilimumab, tremelimumab, nivolumab, pembrolizumab, CT-011, AMP-224, and MDX-1106.

“PD-1 antagonist” or “PD-1 pathway antagonist” means any chemical compound or biological molecule that blocks binding of PD-L1 expressed on a cancer cell to PD-1 expressed on an immune cell (T-cell, B-cell, or NKT-cell) and preferably also blocks binding of PD-L2 expressed on a cancer cell to the immune-cell expressed PD-1. Alternative names or synonyms for PD-1 and its ligands include: PDCD1, PD1, CD279, and SLEB2 for PD-1; PDCDIL1, PDL1, B7H1, B7-4, CD274, and B7-H for PD-L1; and PDCDIL2, PDL2, B7-DC, Btdc, and CD273 for PD-L2. In any of the treatment method, medicaments and uses of the present disclosure in which a human individual is being treated, the PD-1 antagonist blocks binding of human PD-L1 to human PD-1, and preferably blocks binding of both human PD-L1 and PD-L2 to human PD-1. Human PD-1 amino acid sequences can be found in NCBI Locus No.: NP_005009. Human PD-L1 and PD-L2 amino acid sequences can be found in NCBI Locus No.: NP_054862 and NP_079515, respectively.

PD-1 antagonists useful in any of the treatment method, medicaments and uses of the present disclosure include a monoclonal antibody (mAb), or antigen binding fragment thereof, which specifically binds to PD-1 or PD-L1, and preferably specifically binds to human PD-1 or human PD-L1. The mAb may be a human antibody, a humanized antibody, or a chimeric antibody and may include a human constant region. In some embodiments, the human constant region is selected from the group consisting of IgG1, IgG2, IgG3, and IgG4 constant regions, and in preferred embodiments, the human constant region is an IgG1 or IgG4 constant region. In some embodiments, the antigen binding fragment is selected from the group consisting of Fab, Fab′-SH, F(ab′)₂, scFv, and Fv fragments.

Examples of mAbs that bind to human PD-1, and useful in the treatment method, medicaments and uses of the present disclosure, are described in U.S. Pat. Nos. U.S. Pat. Nos. 7,488,802, 7,521,051, 8,008,449, 8,354,509, and 8,168,757, PCT International Patent Application Publication Nos. WO2004/004771, WO2004/072286, and WO2004/056875, and U.S. Patent Application Publication No. US2011/0271358.

Examples of mAbs that bind to human PD-L1, and useful in the treatment method, medicaments and uses of the present disclosure, are described in PCT International Patent Application Nos. WO2013/019906 and WO2010/077634 A1 and in U.S. Pat. No. 8,383,796. Specific anti-human PD-L1 mAbs useful as the PD-1 antagonist in the treatment method, medicaments and uses of the present disclosure include MPDL3280A, BMS-936559, MEDI4736, MSB0010718C, and an antibody that comprises the heavy chain and light chain variable regions of SEQ ID NO:24 and SEQ ID NO:21, respectively, of WO2013/019906.

Other PD-1 antagonists useful in any of the treatment method, medicaments, and uses of the present disclosure include an immune-adhesion that specifically binds to PD-1 or PD-L1, and preferably specifically binds to human PD-1 or human PD-L1, e.g., a fusion protein containing the extracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to a constant region such as an Fc region of an immunoglobulin molecule. Examples of immune-adhesion molecules that specifically bind to PD-1 are described in PCT International Patent Application Publication Nos. WO2010/027827 and WO2011/066342. Specific fusion proteins useful as the PD-1 antagonist in the treatment method, medicaments, and uses of the present disclosure include AMP-224 (also known as B7-DCIg), which is a PD-L2-FC fusion protein and binds to human PD-1.

Thus, the invention further relates to a method of treating cancer in a human patient comprising administration of a compound of Formula Ia and Formula I and a PD-1 antagonist to the patient. The compound of the invention and the PD-1 antagonist may be administered concurrently or sequentially.

In particular embodiments, the PD-1 antagonist is an anti-PD-1 antibody, or antigen binding fragment thereof. In alternative embodiments, the PD-1 antagonist is an anti-PD-L1 antibody, or antigen binding fragment thereof. In some embodiments, the PD-1 antagonist is pembrolizumab (KEYTRUDA™, Merck & Co., Inc., Kenilworth, N.J., USA), nivolumab (OPDIVO™, Bristol-Myers Squibb Company, Princeton, N.J., USA), cemiplimab (LIBTAYO™, Regeneron Pharmaceuticals, Inc., Tarrytown, N.Y., USA), atezolizumab (TECENTRIQ™, Genentech, San Francisco, Calif., USA), durvalumab (IMFINZI™, AstraZeneca Pharmaceuticals LP, Wilmington, Del.), or avelumab (BAVENCIO™, Merck KGaA, Darmstadt, Germany).

In some embodiments, the PD-1 antagonist is pembrolizumab. In particular sub-embodiments, the method comprises administering 200 mg of pembrolizumab to the patient about every three weeks. In other sub-embodiments, the method comprises administering 400 mg of pembrolizumab to the patient about every six weeks.

In further sub-embodiments, the method comprises administering 2 mg/kg of pembrolizumab to the patient about every three weeks. In particular sub-embodiments, the patient is a pediatric patient.

In some embodiments, the PD-1 antagonist is nivolumab. In particular sub-embodiments, the method comprises administering 240 mg of nivolumab to the patient about every two weeks.

In other sub-embodiments, the method comprises administering 480 mg of nivolumab to the patient about every four weeks.

In some embodiments, the PD-1 antagonist is cemiplimab. In particular embodiments, the method comprises administering 350 mg of cemiplimab to the patient about every 3 weeks.

In some embodiments, the PD-1 antagonist is atezolizumab. In particular sub-embodiments, the method comprises administering 1200 mg of atezolizumab to the patient about every three weeks.

In some embodiments, the PD-1 antagonist is durvalumab. In particular sub-embodiments, the method comprises administering 10 mg/kg of durvalumab to the patient about every two weeks.

In some embodiments, the PD-1 antagonist is avelumab. In particular sub-embodiments, the method comprises administering 800 mg of avelumab to the patient about every two weeks.

Examples of other cytotoxic agents include, but are not limited to, arsenic trioxide (sold under the tradename TRISENOX®), asparaginase (also known as L-asparaginase, and Erwinia L-asparaginase, sold under the tradenames ELSPAR® and KIDROLASE®).

Chemotherapeutic agents that may be used in combination with the compounds of general Formula Ia and Formula I disclosed herein include abiraterone acetate, altretamine, anhydrovinblastine, auristatin, bexarotene, bicalutamide, BMS 184476, 2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl)benzene sulfonamide, bleomycin, N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-proly-1-Lproline-tbutylamide, cachectin, cemadotin, chlorambucil, cyclophosphamide, 3′,4′-didehydro-4′deoxy-8′-norvin-caleukoblastine, docetaxol, doxetaxel, cyclophosphamide, carboplatin, carmustine, cisplatin, cryptophycin, cyclophosphamide, cytarabine, dacarbazine (DTIC), dactinomycin, daunorubicin, decitabine dolastatin, doxorubicin (adriamycin), etoposide, 5-fluorouracil, finasteride, flutamide, hydroxyurea and hydroxyureataxanes, ifosfamide, liarozole, lonidamine, lomustine (CCNU), MDV3100, mechlorethamine (nitrogen mustard), melphalan, mivobulin isethionate, rhizoxin, sertenef, streptozocin, mitomycin, methotrexate, taxanes, nilutamide, nivolumab, onapristone, paclitaxel, pembrolizumab, prednimustine, procarbazine, RPR109881, stramustine phosphate, tamoxifen, tasonermin, taxol, tretinoin, vinblastine, vincristine, vindesine sulfate, and vinflunine.

Examples of vascular endothelial growth factor (VEGF) receptor inhibitors include, but are not limited to, bevacizumab (sold under the trademark AVASTIN by Genentech/Roche), axitinib (described in PCT International Patent Publication No. WO01/002369), Brivanib Alaninate ((S)—((R)-1-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-5-methylpyrrolo[2,1-f][1,2,4]triazin-6-yloxy)propan-2-yl)2-aminopropanoate, also known as BMS-582664), motesanib (N-(2,3-dihydro-3,3-dimethyl-TH-indol-6-yl)-2-[(4-pyridinylmethyl)amino]-3-pyridinecarboxamide. and described in PCT International Patent Application Publication No. WO02/068470), pasireotide (also known as SO 230, and described in PCT International Patent Publication No. WO02/010192), and sorafenib (sold under the tradename NEXAVAR).

Examples of topoisomerase II inhibitors, include but are not limited to, etoposide (also known as VP-16 and Etoposide phosphate, sold under the tradenames TOPOSAR, VEPESID, and ETOPOPHOS), and teniposide (also known as VM-26, sold under the tradename VUMON).

Examples of alkylating agents, include but are not limited to, 5-azacytidine (sold under the trade name VIDAZA), decitabine (sold under the trade name of DECOGEN), temozolomide (sold under the trade names TEMODAR and TEMODAL by Schering-Plough/Merck), dactinomycin (also known as actinomycin-D and sold under the tradename COSMEGEN), melphalan (also known as L-PAM, L-sarcolysin, and phenylalanine mustard, sold under the tradename ALKERAN), altretamine (also known as hexamethylmelamine (HMM), sold under the tradename HEXALEN), carmustine (sold under the tradename BCNU), bendamustine (sold under the tradename TREANDA), busulfan (sold under the tradenames BUSULFEX® and MYLERAN®), carboplatin (sold under the tradename PARAPLATIN®), lomustine (also known as CCNU, sold under the tradename CEENU®), cisplatin (also known as CDDP, sold under the tradenames PLATINOL® and PLATINOL®-AQ), chlorambucil (sold under the tradename LEUKERAN®), cyclophosphamide (sold under the tradenames CYTOXAN® and NEOSAR®), dacarbazine (also known as DTIC, DIC and imidazole carboxamide, sold under the tradename DTIC-DOME®), altretamine (also known as hexamethylmelamine (HMM) sold under the tradename HEXALEN®), ifosfamide (sold under the tradename IFEX®), procarbazine (sold under the tradename MATULANE®), mechlorethamine (also known as nitrogen mustard, mustine and mechloroethamine hydrochloride, sold under the tradename MUSTARGEN®), streptozocin (sold under the tradename ZANOSAR®), thiotepa (also known as thiophosphoamide, TESPA and TSPA, and sold under the tradename THIOPLEX®.

Examples of anti-tumor antibiotics include, but are not limited to, doxorubicin (sold under the tradenames ADRIAMYCIN® and RUBEX®), bleomycin (sold under the tradename LENOXANE®), daunorubicin (also known as dauorubicin hydrochloride, daunomycin, and rubidomycin hydrochloride, sold under the tradename CERUBIDINE®), daunorubicin liposomal (daunorubicin citrate liposome, sold under the tradename DAUNOXOME®), mitoxantrone (also known as DHAD, sold under the tradename NOVANTRONE®), epirubicin (sold under the tradename ELLENCE™) idarubicin (sold under the tradenames IDAMYCIN®, IDAMYCIN PFS®), and mitomycin C (sold under the tradename MUTAMYCIN®).

Examples of anti-metabolites include, but are not limited to, claribine (2-chlorodeoxyadenosine, sold under the tradename LEUSTATIN®), 5-fluorouracil (sold under the tradename ADRUCIL®), 6-thioguanine (sold under the tradename PURINETHOL®), pemetrexed (sold under the tradename ALINMTA®), cytarabine (also known as arabinosylcytosine (Ara-C), sold under the tradename CYTOSAR-U®), cytarabine liposomal (also known as Liposomal Ara-C, sold under the tradename DEPOCYT™), decitabine (sold under the tradename DACOGEN®), hydroxyurea (sold under the tradenames HYDREA®, DROXIA™ and MYLOCEL™), fludarabine (sold under the tradename FLUDARA®), floxuridine (sold under the tradename FUDR®), cladribine (also known as 2-chlorodeoxyadenosine (2-CdA) sold under the tradename LEUSTATIN™), methotrexate (also known as amethopterin, methotrexate sodium (MTX), sold under the tradenames RHEUMATREX® and TREXALL™), and pentostatin (sold under the tradename NIPENT®).

Examples of retinoids include, but are not limited to, alitretinoin (sold under the tradename PANRETIN®), tretinoin (all-trans retinoic acid, also known as ATRA, sold under the tradename VESANOID®), isotretinoin (13-c/s-retinoic acid, sold under the tradenames ACCUTANE®, AMNESTEEM®, CLARAVIS®, CLARUS®, DECUTAN®, ISOTANE®, IZOTECH®, ORATANE®, ISOTRET®, and SOTRET®), and bexarotene (sold under the tradename TARGRETIN®).

Further disclosed herein is a compound disclosed herein, or a pharmaceutically acceptable salt thereof, for use in therapy. In one embodiment, disclosed herein is the use of a compound disclosed herein, or a pharmaceutically acceptable salt, solvate or hydrate thereof, for the preparation of a medicament for use in therapy.

Intermediates

Intermediates were either purchased from the following vendors or synthesized using the cited literature references as shown or further modified through additional synthetic manipulations analogous to phosphoramidites described in intermediates I-1 to I-21.

TABLE 1 Intermediates used in the synthesis of Examples of the instant invention. Structure Bases Source I-1

B = nucleobase A, C, G, U ChemGenes Vendor I-2

B = nucleobase A, C, G, U ChemGenes Vendor I-3

B = nucleobase A, C, G, U ChemGenes Vendor I-4

B = nucleobase A, C, G, U ChemGenes Vendor I-5

B = nucleobase A, C, G, U Glen Research Vendor I-6

B = nucleobase A, 5-Me-C, G, T Exiqon Vendor I-7

B = nucleobase A, 5-Me-C, G, T ChemGenes Vendor I-8

B = nucleobase A, C, G, U J . Med. Chem. 2000, 43, 4516-4525 I-9

B = nucleobase A, C, G, U Canadian Journal of Chemistry 1989, 67, 831-9 I-11

B = nucleobase A, C, G, U ChemGenes Vendor I-12

B = nucleobase A, C, G, U J . Med. Chem. 2006, 49, 1624-1634 I-13

B = nucleobase A, C, G, U Nucleic Acids Research, 2009, Vol. 37, No. 4 1353-1362 I-14

B = nucleobase A, C, G, U Nuc. Acids Res. 2015, 43, 7189-7200. I-15

B = nucleobase A, C, G, U U.S., 4,849,513, 18 Jul. 1989; Nucleosides & Nucleotides 1992, 11, 1333-51 I-17

Journal of the Chemical Society, Perkin Transactions 1: Organic and Bio- Organic Chemistry 1995 (12), 1543-50 I-18

Carbosynth Vendor I-19

Carbosynth Vendor I-20

Barry and Associates Vendor I-21

WO/2017/027646 I-22

Carbosynth Vendor I-23

Journal of Organic Chemistry, 2013, 78, 11271-11282. I-24

Barry and Associates Vendor I-25

Barry and Associates Vendor

The following experimental procedures detail the preparation of intermediates used in the synthesis of the Examples of the instant invention. The exemplified procedures are for illustrative purposes only, and are not intended to limit the scope of the instant invention in any way.

Step 1: To a solution of N-(1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)-tetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)acetamide (200 g, 701.13 mmol, 1.00 equiv) in pyridine (3000 mL) was added the solution of 1,3-dichloro-1,1,3,3-teraisopropyldisiloxane (TIPSCl₂) (240 g, 762 mmol, 1.1 equiv) in pyridine dropwise. The resulting solution was allowed to react overnight with stirring at room temperature. The reaction mixture was concentrated under vacuum and the residue was extracted with EtOAc three times. The combined organic layer was washed with brine twice and dried over anhydrous Na₂SO₄ The filtrate was concentrated under vacuum and the residue was purified by a silica gel column, eluting with dichloromethane/methanol (100:1) to afford of N-(1-((6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetraisopropyl-tetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)acetamide. LCMS-(ES, m/z) found: 528[M+H]⁺. ¹H NMR (300 MHz, Chloroform-d) δ 9.99 (s, 1H), 8.19 (d, J=7.5 Hz, 1H), 7.43 (d, J=7.4 Hz, 1H), 5.80 (s, 1H), 4.32-4.15 (m, 4H), 4.00 (dd, J=13.4, 2.3 Hz, 1H), 3.34 (s, 1H), 2.28 (s, 3H), 1.14-0.86 (m, 28H).

Step 2: To a solution of N-(1-((6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)acetamide (350 g, 663 mmol, 1.00 equiv) in dichloromethane (1500 mL) was added methyl 1H-1,2,4-triazole-1-carbodithioate (126 g, 794 mmol, 1.20 equiv) in several batches. To the above was added 1,8-diazabicyclo[5.4.0]-undec-7-ene (DBU) (152 g, 996 mmol, 1.50 equiv) dropwise with stirring at 0° C., and stirred overnight at room temperature. The resulting mixture was neutralized with hydrochloric acid (2 M) and the organic layer was washed with sodium bicarbonate solution (sat.), then dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column and eluted with dichloromethane/methanol (100:1) to afford 0-((6aR,8R,9R,9aR)-8-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-9-yl) S-methyl carbonodithioate.

LCMS (ES, m/z) found: 618 [M+H]⁺. ¹H NMR (300 MHz, DMSO-d₆) δ 10.91 (s, 1H), 8.05 (d, J=7.5 Hz, 1H), 7.20 (d, J=7.5 Hz, 1H), 6.34 (dd, J=5.3, 1.1 Hz, 1H), 5.80 (d, J=1.1 Hz, 1H), 4.70 (dd, J=8.5, 5.4 Hz, 1H), 4.07-3.91 (m, 3H), 2.58 (s, 3H), 2.09 (s, 3H), 1.07-0.77 (m, 28H).

Step 3: To a solution of 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (23 g, 80.44 mmol, 5.00 equiv) in dichloromethane (230 mL) was added HF/Py (105 mL, 70% HF in Py) dropwise with stirring at −78° C. The mixture was stirred for 10 minutes at −78° C. To this was added a solution of N-[1-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide (10 g, 16.18 mmol, 1.00 equiv) in dichloromethane (70 mL) dropwise with stirring at −78° C. The resulting solution was stirred for 3 h at 0° C. in an ice/salt bath. The reaction was then quenched by the addition of 100 mL of NaHSO₃ (1M). The pH value of the solution was adjusted to 8 with sodium bicarbonate (solid). The solids were filtered out. The resulting solution was extracted with 4×300 mL of ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with dichloromethane/methanol/TEA (95:5:0.3) as eluent to furnish 800 mg (13%) of N-[1-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide as a white solid. The above procedure was repeated for ten batches (20 g×10), and the crude was purified together to give N-(1-((2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)tetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)acetamide.

LCMS (ES, m/z) found: 354 [M+H]⁺. ¹H NMR (300 MHz, DMSO-d6) δ 10.92 (s, 1H), 8.38 (d, J=7.5 Hz, 1H), 7.18 (d, J=7.5 Hz, 1H), 6.00 (d, J=3.4 Hz, 1H), 5.76 (d, J=6.0 Hz, 1H), 5.28 (t, J=4.9 Hz, 1H), 4.97-4.81 (m, 1H), 4.21 (p, J=5.4 Hz, 1H), 3.92 (dt, J=6.1, 2.9 Hz, 1H), 3.81-3.68 (m, 1H), 3.65-3.52 (m, 1H), 2.07 (s, 3H).

Step 4: To a solution of N-[1-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide (9 g, 25.48 mmol, 1.00 equiv, co-evaporated with anhydrous pyridine three times) in pyridine (100 mL) was added a solution of 1-[chloro(4-methoxyphenyl)benzyl]-4-methoxybenzene (17.1 g, 50.47 mmol, 2.00 equiv, co-evaporated with anhydrous pyridine three times) in pyridine (200 mL) dropwise with stirring at 30° C. After the resulting solution was stirred for 3 h at 30° C., then concentrated under vacuum. The resulting solution was diluted with 400 mL of DCM, then was washed with 1×300 mL of sodium bicarbonate (sat.) and 1×300 mL of brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with ethyl acetate/petroleum ether/Et₃N (60:40:0.3) as eluent to furnish N-[1-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide. LCMS (ES, m/z) found: 656 [M+H]⁺. ¹H NMR (400 MHz, Chloroform-d) δ 9.49 (s, 1H), 8.35 (d, J=7.5 Hz, 1H), 7.43 (d, J=7.7 Hz, 2H), 7.39-7.26 (m, 6H), 7.25-7.07 (m, 2H), 6.88 (d, J=8.5 Hz, 4H), 6.16 (s, 1H), 5.16-5.10 (m, 1H), 4.60 (t, J=6.0 Hz, 1H), 4.24 (d, J=7.5 Hz, 1H), 3.83 (s, 6H), 3.65 (d, J=11.3 Hz, 1H), 3.58 (d, J=11.2 Hz, 1H), 3.34 (s, 1H), 2.22 (s, 3H).

Step 5: To a solution of N-[1-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide (13.0 g, 19.83 mmol, 1.00 equiv, which dried by co-evaporation with pyridine twice) in dichloromethane (200 mL) was added DIPEA (12.8 g, 99.04 mmol, 5.00 equiv) dropwise with stirring at 0° C. To the mixture was added 3-([bis(propan-2-yl)amino](chloro)phosphanyloxy)propanenitrile (7.0 g, 29.58 mmol, 1.50 equiv) dropwise with stirring at 0° C. After the resulting solution was stirred for 2 h at room temperature, it then was diluted with 500 mL of CH₂Cl₂. The resulting mixture was washed with 2×300 mL of saturated sodium bicarbonate and 300 mL of brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with petroleum ether (PE):ethyl acetate (EA):TEA (38:62:0.5) as eluent to obtain the crude. The crude product (13 g) was purified by Flash-Prep-HPLC with the following conditions (IntelFlash-1): Column, C18 silica gel; mobile phase, 50% CH₃CN/H₂O (0.5% NH₄HCO₃) increasing to 80% CH₃CN/H₂O (0.5% NH₄HCO₃) over 30 min; Detector, UV 254 nm. This resulted in N-[1-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-([[bis(propan-2-yl)amino](2-cyanoethoxy)phosphanyl]oxy)-3-(trifluoromethoxy)oxolan-2-yl]-2-oxo-1,2-dihydropyrimidin-4-yl]acetamide. L-MS (ES, m/z) found: 856 [M+H]⁺. ¹H-NMR (300 MHz, Chloroform-d,ppm): δ 10.69-10.57 (m, 1H), 8.30 (m, 1H), 7.43 (m, 2H), 7.39-7.23 (m, 7H), 7.11 (m, 1H), 6.89 (m, 4H), 6.23 (d, J=2.8 Hz, 1H), 4.98 (m, 1H), 4.66 (m, 1H), 4.40-4.23 (m, 1H), 3.98-3.45 (m, 12H), 2.62 (t, J=6.2 Hz, 1H), 2.43 (m, 1H), 2.28 (d, J=1.8 Hz, 3H), 1.24-1.11 (m, 9H), 1.08 (d, J=6.8 Hz, 3H). F-NMR: (300 MHz, Chloroform-d, ppm): δ 58.148, −58.222. P-NMR: (300 MHz, Chloroform-d, ppm): δ 15.8, 150.9.

Step 1: I-23C was made in accordance to the procedure described in the procedure for making Intermediate 1-23. To a solution of O-(6aR,8R,9aR)-8-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2,2,4,4-tetraisopropyl-tetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-9-yl S-methyl carbonodithioate (270 g, 425 mmol, 1.00 equiv) in acetic anhydride (1000 mL) and AcOH (1000 mL) was added sulfuric acid (10 mL) dropwise with stirring at 0° C. under inert atmosphere. The resulting solution was allowed to stir overnight at room temperature. The solvent was concentrated under vacuum and the residue was extracted with EtOAc three times. The combined organic phase was washed with NaHCO₃(sat.) twice and brine once, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column, eluting with dichloromethane/methanol (100:1) and further purification by recrystallization from acetonitrile:ether=1:10 to afford (2R,3R,4R,5R)-5-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2-(acetoxymethyl)-4 (((methylthio)carbonothioyl)oxy)tetrahydrofuran-3-yl acetate.

LCMS:(ES, m/z) found:460 [M+H]⁺. ¹H NMR (400 MHz, Chloroform-d) δ 10.47 (s, 1H), 7.80 (d, J=7.5 Hz, 1H), 7.50 (d, J=7.6 Hz, 1H), 6.30 (dd, J=5.9, 3.7 Hz, 1H), 6.03 (d, J=3.7 Hz, 1H), 5.53 (t, J=6.1 Hz, 1H), 4.51-4.38 (m, 3H), 2.58 (s, 3H), 2.28 (s, 3H), 2.15 (s, 3H), 2.10 (s, 3H).

Step 2: To a solution of 1,3-dibromo-5-isopropylimidazolidine-2,4-dione (47 g, 165 mmol, 5 equiv) in dichloromethane (300 mL) was added HF/Py (150 mL, 70% HF in Py) dropwise with stirring at −78° C. under inert atmosphere and kept stirring for 10 min. This was followed by the addition of a solution of (2R,3R,4R,5R)-5-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2-(acetoxymethyl)-4-(((methylthio)carbonothi-oyl)oxy)tetrahydrofuran-3-yl acetate (15 g, 32.68 mmol, 1.00 equiv) in dichloromethane (200 mL) dropwise with stirring at −78° C., and then kept stirring for 2 h at −20° C. The resulting mixture was quenched by the addition of NaHSO₃ (sat.) and extracted with dichloromethane four times. The combined organic layers was washed with sodium bicarbonate (sat.) twice and brine once, and dried over anhydrous sodium sulfate. The filtrate was concentrated and residue was purified by recrystallization from ether to afford (2R,3R,4R,5R)-5-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2-(acetoxymethyl)-4-(trifluoromethoxy)tetrahydrofuran-3-yl acetate. LCMS (ES, m/z) found: 438 [M+H]⁺. ¹H NMR (300 MHz, Chloroform-d) δ 10.13 (s, 1H), 7.91 (d, J=7.6 Hz, 1H), 7.46 (d, J=7.6 Hz, 1H), 5.91 (d, J=1.5 Hz, 1H), 5.16 (d, J=5.9 Hz, 2H), 4.57-4.17 (m, 3H), 2.24 (s, 3H), 2.16-1.97 (m, 6H).

Step 3: To a solution of 2R,3R,4R,5R)-5-(4-acetamido-2-oxopyrimidin-1(2H)-yl)-2-(acetoxymethyl)-4-(trifluoromethoxy)tetrahydrofuran-3-yl acetate (38 g, 87 mmol, 1.00 equiv) in MeOH (260 mL) was added NH₃ (130 mL, 7 M in MeOH) dropwise at 0° C. The resulting solution was stirred overnight at room temperature and then concentrated under vacuum to give 4-amino-1-((2R, 3R, 4R, 5R)-4-hydroxy-5-(hydro-xylmethyl)-3-(trifluoromethoxy)-tetrahydrofuran-2-yl) pyrimidin-2(1H)-one. LCMS: (ES, m/z): 312 [M+H]⁺.

Step 4: To a solution of crude 4-amino-1-((2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluorometh-oxy)-tetrahydrofuran-2-yl)pyrimidin-2(1H)-one (15 g, 48 mmol, 1.00 equiv) in water (100 mL) was added NaNO₂ (67 g, 971 mmol, 20 equiv) in several batches, and followed added acetic acid (50 mL) dropwise at 0° C. The resulting solution was stirred overnight at room temperature and concentrated under vacuum. The residue was triturated with EtOH and the filtrate was concentrated under vacuum after filtration to afford 1-((2R, 3R, 4R, 5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)-tetrahydrofuran-2-yl) pyrimidine-2,4(1H, 3H)-dione. LCMS (ES, m/z): 313[M+H]⁺.

Step 5: To a solution of 1-((2R, 3R, 4R, 5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)-tetrahydrofuran-2-yl) pyrimidine-2, 4(1H, 3H)-dione (14 g, 45 mmol, 1.00 equiv, coevaporate with pyridine twice previously) in pyridine (250 mL) was added a solution of 1-(chlorobis(4-methoxyphenyl) methyl)benzene (23 g, 67 mmol, 1.50 equiv) in pyridine (50 mL) dropwise with stirring at room temperature under inert atmosphere. The resulting solution was stirred overnight at room temperature and then concentrated under vacuum. The residue was extracted with EtOAc twice and the combined organic layer was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column, eluting with dichloromethane/methanol/Et₃N (100:1:0.5) to afford 1-((2R, 3R, 4R, 5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-hydroxy-3-(trifluoromethoxy)-tetrahydrofuran-2-yl) pyrimidine-2, 4(1H, 3H)-dione. ¹H NMR (300 MHz, Chloroform-d) δ 8.75 (d, J=2.0 Hz, 1H), 7.68 (d, J=8.2 Hz, 1H), 7.36-7.17 (m, 9H), 6.90-6.77 (m, 4H), 6.21 (d, J=5.2 Hz, 1H), 5.30 (dd, J=8.2, 2.2 Hz, 1H), 4.91 (t, J=5.0 Hz, 1H), 4.53 (q, J=4.5 Hz, 1H), 4.18-4.11 (m, 2H), 3.77 (s, 6H), 3.54-3.50 (m, 1H), 2.50 (d, J=5.0 Hz, 1H).

Step 6: 1-((2R, 3R, 4R, 5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-hydroxy-3-(trifluoro-methoxy)-tetrahydrofuran-2-yl) pyrimidine-2, 4(1H, 3H)-dione (20 g, 33 mmol, 1.00 equiv) was co-evaporated with dry pyridine three times and then diluted with dry dichloromethane (200 mL), To the above solution was added N-ethyl-N-isopropylpropan-2-amine (13 g, 99 mmol, 3.00 equiv), followed by addition of 3-chloro-3-(diisopropylamino) phosphinooxypropanenitrile (12 g, 50 mmol, 1.50 equiv) dropwise with stirring at room temperature under inert atmosphere. The resulting solution was stirred for 2 h at room temperature and then quenched with sodium bicarbonate (sat.) and extracted with dichloromethane twice. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column, eluting with ethyl acetate/hexane/EtN₃ (50:100:0.75) and then further purified by a reverse column, eluting with acetonitrile: water (containing 0.1% NH₄HCO₃) from 20:80 to 80:20 over 1.5 h to afford (2R,3R,4R,5R)-2-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-(trifluoromethoxy)tetrahydrofuran-3-yl (2-cyanoethyl) diisopropylphosphoramidite. LCMS (ES, m/z) found: 815 [M+H]⁺. ¹H-NMR (400 MHz, CDCl₃, ppm): 8.82 (s, 1H), 7.76 (dd, J=11.9, 8.1 Hz, 1H), 7.42-7.25 (m, 7H), 6.88 (dd, J=8.6, 3.8 Hz, 4H), 6.23 (dd, J=9.3, 4.9 Hz, 1H), 5.33 (dd, J=8.3, 4.2 Hz, 1H), 5.01 (q, J=5.7 Hz, 1H), 4.69 (ddt, J=14.7, 9.4, 4.7 Hz, 1H), 4.34 (d, J=3.5 Hz, 1H), 4.23 (d, J=4.5 Hz, 0H), 3.93 (dt, J=13.4, 6.4 Hz, 0H), 3.86-3.72 (m, 6H), 3.65 (dq, J=12.3, 6.3, 5.8 Hz, 3H), 3.61-3.44 (m, 2H), 2.66 (t, J=6.2 Hz, 1H), 2.48 (s, 1H), 1.21 (t, J=5.5 Hz, 10H), 1.10 (d, J=6.7 Hz, 2H). ³¹P NMR-(400 MHz, CDCl₃, ppm): 151.5, 152.0. ¹⁹F NMR: (400 MHz, CDCl₃, ppm): δ−58.6, −59.1.

Step 1: To a solution of 1H-1,2,4-triazole (250 g, 3.62 mol, 1.00 equiv) in acetone (7000 mL) was added K₃PO₄ (768 g, 3.62 mol, 1.00 equiv). After the mixture was stirred for 20 minutes at 25° C., then was added CS₂ (826 g, 3.00 equiv) dropwise with stirring at 25° C. After the mixture was stirred for 20 minutes at 25° C., then was added CH₃I (515 g, 3.63 mol, 1.00 equiv) dropwise with stirring at 25° C. After the resulting solution was stirred overnight at 25° C., the solids were filtered out. The filtrate was concentrated under vacuum, then was dissolved in 5 L of ethyl acetate. The insoluble solids were filtered out. The filtrate was concentrated under vacuum to get the crude. The crude product was re-crystallized from EA:Et₂O:PE with the ratio of 1:1:1 to afford 1-(methylsulfanyl)carbothioyl-1H-1,2,4-triazole. LCMS (ES, m/z) found: 160[M+H]⁺. ¹H NMR (300 MHz, Chloroform-d) δ 9.14 (s, 1H), 8.06 (s, 1H), 2.72 (s, 3H).

Step 2: To a solution of N-[9-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-9H-purin-6-yl]benzamide (200 g, 538.58 mmol, 1.00 equiv) in anhydrous pyridine (2500 mL) was added (dichloro-4-sulfanylidene)(titanio)phosphane (186 g, 1.02 mol, 1.11 equiv) dropwise with stirring at room temperature. After the resulting solution was stirred overnight at room temperature, the crude mixture was concentrated under vacuum. The residue was dissolved in 1000 mL of dichloromethane, then was washed with 2×400 mL of sodium bicarbonate (sat.), dried over anhydrous sodium sulfate. The residue was purified by a silica gel column with ethyl acetate/petroleum ether (2:1) as eluent to N-[9-[(6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-9H-purin-6-yl]benzamide. LCMS (ES, m/z) found: 614 [M+H]⁺. ¹H NMR (300 MHz, DMSO-d₆) δ 11.17 (s, 1H), 8.63 (s, 1H), 8.50 (s, 1H), 8.05-7.96 (m, 2H), 7.61 (t, J=7.3 Hz, 1H), 7.52 (dd, J=8.3, 6.7 Hz, 2H), 5.97 (d, J=1.4 Hz, 1H), 5.64 (d, J=4.7 Hz, 1H), 4.79 (dd, J=8.3, 5.1 Hz, 1H), 4.62 (t, J=5.3 Hz, 1H), 4.10-3.94 (m, 2H), 3.97-3.86 (m, 1H), 1.07-0.88 (m, 28H).

Step 3: To a solution of N-[9-[(6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-9H-purin-6-yl]benzamide (246 g, 400.75 mmol, 1.00 equiv) in dichloromethane (600 mL) was added 1-(methylsulfanyl)carbothioyl-1H-1,2,4-triazole (115 g, 722.21 mmol, 1.80 equiv). This was followed by the addition of DBU (80 g, 525.49 mmol, 1.30 equiv) dropwise with stirring at 25° C. After the resulting solution was stirred for overnight at 25° C., then was washed with 2×300 mL of hydrogen chloride (1 M) and 1×300 mL of sodium bicarbonate (sat.), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with ethyl acetate/petroleum ether (1:1) as eluent to furnish N-[9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-9H-purin-6-yl]benzamide. LCMS (ES, m/z) found: 704[M+H]⁺.

¹H NMR (400 MHz, DMSO-d₆) δ 11.28 (s, 1H), 8.64 (d, J=18.1 Hz, 2H), 8.09-8.02 (m, 2H), 7.67 (t, J=7.4 Hz, 1H), 7.57 (t, J=7.6 Hz, 2H), 6.77 (dd, J=5.7, 1.4 Hz, 1H), 6.41 (d, J=1.4 Hz, 1H), 5.45 (dd, J=8.4, 5.8 Hz, 1H), 4.05 (ddd, J=14.2, 11.1, 6.9 Hz, 3H), 2.65 (s, 3H), 1.28-0.74 (m, 28H).

Step 4: To a solution of 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (20.5 g, 71.70 mmol, 5.00 equiv) in dichloromethane (150 mL) was added a solution of 70% HF in Py (60 mL) dropwise with stirring at −78° C. The mixture was stirred for 10 minutes at −78° C., then added a solution of N-[9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-9H-purin-6-yl]benzamide (10 g, 14.20 mmol, 1.00 equiv) in dichloromethane (50 mL) dropwise with stirring at −78° C. The mixture was stirred for 10 minutes at −78° C. followed by stirring for 3 h at −5° C. The reaction was then quenched by the addition of 10 g of NaHSO₃. The pH value of the solution was adjusted to 7-8 with sodium bicarbonate (sat.). The solids were filtered out. The filtrate was extracted with 2×200 mL of dichloromethane and the combined organic layers were dried over anhydrous sodium sulfate. The residue was purified by a silica gel column with dichloromethane/methanol (25:1) as eluent to furnish N-[9-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-9H-purin-6-yl]benzamide. LCMS (ES, m/z) found: 440 [M+H]⁺. ¹H-NMR: (300 MHz, CDCl₃, ppm): δ 8.81 (s, 1H), 8.14-7.98 (m, 3H), 7.70-7.48 (m, 3H), 6.14 (d, J=7.4 Hz, 1H), 5.62 (dd, J=7.5, 4.4 Hz, 1H), 4.72 (d, J=4.4 Hz, 1H), 4.40 (s, 1H), 3.99 (dd, J=13.2, 1.6 Hz, 1H), 3.84-3.68 (m, 1H), 3.48 (s, 2H).

Step 5: N-[9-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-9H-purin-6-yl]benzamide (11 g, 25.04 mmol, 1.00 equiv) was co-evaporated with anhydrous pyridine three times before added into a 500-mL 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen. This was followed by the addition of anhydrous pyridine (110 mL) and a solution of 1-[chloro(4-methoxyphenyl)benzyl]-4-methoxybenzene (17 g, 50.17 mmol, 2.00 equiv, was dried by co-evaporation with anhydrous pyridine three times) in anhydrous pyridine (170 mL) dropwise with stirring at 30° C. The resulting solution was stirred for 3 h at 30° C., then was concentrated under vacuum. The resulting solution was diluted with 500 mL of dichloromethane, then was washed with 2×200 mL of sodium bicarbonate (sat.) and 1×200 mL of brine (sat.), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with PE:EA:TEA (50:50:0.3) as eluent to furnish N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-9H-purin-6-yl]benzamide. LCMS (ES, m/z) found: 742 [M+H]⁺.

¹H NMR (400 MHz, Chloroform-d) δ 9.06 (s, 1H), 8.69 (s, 1H), 8.13 (s, 1H), 8.04 (d, J=8.0 Hz, 2H), 7.63 (t, J=7.3 Hz, 1H), 7.54 (t, J=7.7 Hz, 2H), 7.46-7.39 (m, 2H), 7.35-7.19 (m, 5H), 7.19 (d, J=8.6 Hz, 1H), 6.84 (t, J=9.1 Hz, 4H), 6.33 (d, J=5.9 Hz, 1H), 5.83 (t, J=5.4 Hz, 1H), 4.75 (dd, J=10.1, 4.9 Hz, 1H), 4.32 (q, J=3.6 Hz, 1H), 3.81 (d, J=8.2 Hz, 6H), 3.58 (dd, J=10.7, 3.6 Hz, 1H), 3.44 (dd, J=10.8, 3.8 Hz, 1H), 3.13-3.03 (m, 1H).

Step 6: N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-9H-purin-6-yl]benzamide (13.5 g, 18.20 mmol, 1.00 equiv) was co-evaporated with anhydrous pyridine three times before added into a 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen. This was followed by the addition of dichloromethane (140 mL) and DIPEA (12 g, 92.85 mmol, 5.00 equiv). This was followed by the addition of 3-([bis(propan-2-yl)amino](chloro)phosphanyloxy)-propanenitrile (6.5 g, 27.5 mmol, 1.50 equiv) dropwise with stirring at 20° C. The resulting solution was stirred for 1 h at 20° C., then quenched by 5 mL of methanol. The resulting solution was diluted with 500 mL of EA, then was washed with 1×200 mL of sodium bicarbonate (sat.) and 1×200 mL of brine (sat.), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by a silica gel column with PE:EA:TEA (50:50:0.3) as eluent to furnish a crude. The crude was purified by a C18 silica gel column with H₂O(0.5% NH₄HCO₃):ACN(1:99). This resulted in N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-([[bis(propan-2-yl)amino](2-cyanoethoxy)phosphanyl]oxy)-3-(trifluoromethoxy)oxolan-2-yl]-9H-purin-6-yl]benzamide. LCMS (ES, m/z) found: 942 [M+H]⁺. ¹H-NMR: (300 MHz, CDCl₃, ppm): δ 8.97 (d, J=4.9 Hz, 1H), 8.65 (d, J=1.6 Hz, 1H), 8.12 (d, J=4.3 Hz, 1H), 8.04-7.94 (m, 2H), 7.67-7.44 (m, 3H), 7.39 (dt, J=8.0, 1.7 Hz, 2H), 7.32-7.10 (m, 7H), 6.84-6.71 (m, 4H), 6.26 (dd, J=16.3, 6.1 Hz, 1H), 5.90-5.78 (m, 1H), 4.79 (dddd, J=17.7, 12.7, 7.2, 3.7 Hz, 1H), 4.34 (q, J=3.5 Hz, 1H), 3.86 (ddd, J=12.3, 6.0, 1.4 Hz, 1H), 3.75 (d, J=2.4 Hz, 6H), 3.60 (dddd, J=20.4, 13.7, 8.5, 3.9 Hz, 4H), 3.34 (ddd, J=10.7, 6.9, 3.8 Hz, 1H), 2.60 (t, J=6.3 Hz, 1H), 2.39 (td, J=6.4, 2.1 Hz, 1H), 1.19 (dd, J=7.0, 5.7 Hz, 8H), 1.06 (d, J=6.7 Hz, 4H). ³¹PNMR (400 MHz, CDCl₃, ppm): 152.1, 151.9. ¹⁹FNMR (400 MHz, CDCl₃, ppm): δ−59.0, 59.2.

Step 1: To a solution of 2-amino-9-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6,9-dihydro-1H-purin-6-one (100 g, 353.06 mmol, 1.00 equiv) in N,N-dimethylformamide (1000 mL) was added imidazole (88.8 g, 1.31 mol, 3.70 equiv). This was followed by the addition of TiPSCl₂ (166.9 g, 529.84 mmol, 1.50 equiv) dropwise with stirring at room temperature. After the resulting solution was stirred for 2.5 h at room temperature, then quenched by the addition of 1500 mL of water/ice. The solids were collected by filtration and washed with water, then dried in an oven at 50° C. This resulted in (crude) of 9-[(6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f] [1,3, 5,2,4] trioxadisilicon-8-yl]-2-amino-6,9-dihydro-1H-purin-6-one. LC-MS (ES, m/z) found: 526 [M+H]⁺ ¹H NMR (300 MHz, DMSO-d6, ppm) δ 10.61 (s, 1H), 7.73 (s, 1H), 6.45 (s, 2H), 5.64 (d, J=1.8 Hz, 1H), 5.56 (s, 1H), 4.37-4.18 (m, 2H), 4.11-3.83 (m, 3H), 1.05-0.86 (m, 28H).

Step 2: To a solution of 9-[(6aR,8R,9R,9aS)-9-hydroxy-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo [3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-2-amino-6,9-dihydro-1H-purin-6-one (182 g, crude, 346.17 mmol, 1.00 equiv) in dichloromethane (1500 mL), was added 1-(methylsulfanyl)carbothioyl-1H-1,2,4-triazole (71.5 g, 449.03 mmol, 1.20 equiv) under atmosphere of argon. To this was added DBU (79 g, 518.92 mmol, 1.50 equiv) dropwise with stirring at room temperature. After the resulting solution was stirred for 3 h at room temperature, then washed with 2×1000 mL of brine (sat.) and concentrated under vacuum. The residue was purified by a silica gel column with dichloromethane/methanol (30:1) as eluent to furnish 9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis(propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4] trioxadisilicon-8-yl]-2-amino-6,9-dihydro-TH-purin-6-one. LC-MS (ES, m/z) found: 616 [M+H]⁺. ¹H NMR (300 MHz, DMSO-d6) δ 10.69 (s, 1H), 7.88 (s, 1H), 6.38 (dd, J=5.9, 2.7 Hz, 1H), 6.36 (s, 2H), 5.98 (d, J=2.6 Hz, 1H), 4.85-4.74 (m, 1H), 3.99 (tdt, J=12.3, 8.7, 4.1 Hz, 3H), 2.56 (s, 3H), 1.14-0.73 (m, 28H).

Step 3: To a solution of 9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis (propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-2-amino-6,9-dihydro-1H-purin-6-one (113 g, 183.47 mmol, 1.00 equiv) in pyridine (1200 mL), was added TMSCl (100 g, 920.47 mmol, 5.00 equiv) dropwise with stirring at 0° C. and stirred for 2 hours at room temperature. To this was added i-Bu₂O (49.3 g, 312.03 mmol, 1.70 equiv) dropwise with stirring at 0° C. After the resulting solution was stirred for 1 h at room temperature, it then was quenched by the addition of 150 mL saturated aqueous of sodium bicarbonate. The pyridine was evaporated out under vacuum, then diluted with 700 mL of brine (sat.), and extracted with 2 x 800 mL of ethyl acetate. The combined organic layers were concentrated and purified by a silica gel column with dichloromethane/methanol (100% DCM) as eluent to furnish N-[9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis (propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylpropanamide. LCMS (ES, m/z) found: 686 [M+H]⁺

Step 4: To a solution of N-[9-[(6aR,8R,9R,9aR)-9-[[(methylsulfanyl)methanethioyl]oxy]-2,2,4,4-tetrakis (propan-2-yl)-hexahydro-2H-furo[3,2-f][1,3,5,2,4]trioxadisilicon-8-yl]-6-oxo-6,9-dihydro-TH-purin-2-yl]-2-methylpropanamide (90 g, 131.19 mmol, 1.00 equiv) in acetic anhydride (420 mL) and HOAc (210 mL), was added sulfuric acid (conc.) (25.2 g, 256.93 mmol, 1.96 equiv) dropwise with stirring at 0° C. After the resulting solution was stirred overnight at room temperature, then diluted with 700 mL of brine (sat.). The pH value of the solution was adjusted to 7 with a solution of sodium carbonate (4 M), then extracted with dichloromethane 4×1000 ml and the combined organic layer were concentrated under vacuum. The residue was purified by a silica gel column with ethyl acetate/petroleum ether (up to 100% EA) as eluent to furnish [(2R,3R,4R,5R)-3-(acetyloxy)-5-[2-(2-methylpropanamido)-6-oxo-6,9-dihydro-1H-purin-9-yl]-4-[[(methylsulfanyl) methane thioyl]oxy] oxolan-2-yl]methyl acetate. LCMS (ES, m/z) found: 528 [M+H]⁺

Step 5: To a solution of 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (27.1 g, 94.78 mmol, 5.00 equiv) in dichloromethane (140 mL), was added a solution of 70% HF in Py (100 mL) dropwise with stirring at −78° C. To this was added [(2R,3R,4R,5R)-3-(acetyloxy)-5-[2-(2-methylpropanamido)-6-oxo-6,9-dihydro-1H-purin-9-yl]-4-[[(methylsulfanyl)methanethioyl]oxy]oxolan-2-yl]methyl acetate (10 g, 18.95 mmol, 1.00 equiv) with stirring at −78° C. After the resulting solution was stirred for 1 h at −78° C., then quenched by the addition of 100 mL saturated aqueous of NaHSO₃. The resulting solution was extracted with 3×200 mL of ethyl acetate and the organic layers combined. The pH value of the organic layers was adjusted to 8 with a saturated solution of sodium bicarbonate. Then the aqueous phase was extracted with ethyl acetate, and the combined organic layers were concentrated under vacuum. The residue was purified by a silica gel column with ethyl acetate/petroleum ether (up to 100% EA) as eluent to furnish [(2R,3R,4R,5R)-3-(acetyloxy)-5-[2-(2-methylpropanamido)-6-oxo-6,9-dihydro-1H-purin-9-yl]-4-(trifluoromethoxy)oxolan-2-yl]methyl acetate. LCMS (ES, m/z) found: 506 [M+H]⁺.

Step 6: To a solution of guanidine nitrate (11.81 g, 96.84 mmol, 5 equiv), MeONa (1.88 g, 34.86 mmol, 1.80 equiv) in dichloromethane (100 mL) and methanol (900 mL), was added [(2R,3R,4R,5R)-3-(acetyloxy)-5-[2-(2-methylpropanamido)-6-oxo-6,9-dihydro-1H-purin-9-yl]-4-(trifluoromethoxy)oxolan-2-yl]methyl acetate (9.8 g, 19.39 mmol, 1.00 equiv) with stirring at −5° C. After the resulting solution was stirred for 30 min at −5° C., the pH was adjusted 7 with citric acid (1 M) and concentrated under vacuum. The residue was purified by a silica gel column with dichloromethane/methanol (10:1) as eluent to furnish N-[9-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylpropanamide. LCMS (ES, m/z) found: 422 [M+H]⁺.

Step 7: To a solution of N-[9-[(2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-(trifluoromethoxy)oxolan-2-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylpropanamide (7.15 g, 16.97 mmol, 1.00 equiv) in pyridine (70 mL), was added 1-methoxy-4-[1-(4-methoxyphenyl)-1-phenylethyl]benzene (8.63 g, 27.10 mmol, 1.50 equiv) at room temperature. After the resulting solution was stirred for 1.5 h at room temperature, it was concentrated under vacuum. The residue was purified by a silica gel column with EA/PE (up to 80% EA) as eluent to furnish N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl) methoxy] methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylprop anamide. LCMS: (ES, m/z) found: 724 [M+H]⁺.

Step 8: To a solution of N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl)(phenyl)methoxy]methyl]-4-hydroxy-3-(trifluoromethoxy)oxolan-2-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylpropanamide (11.1 g, 15.34 mmol, 1.00 equiv) in dichloromethane (110 mL), was added DIPEA (9.9 g, 76.74 mmol, 5.00 equiv) with stirring at room temperature. To this was added 3-([[bis(propan-2-yl)amino](chloro) phosphanyl]oxy)propanenitrile (7.25 g, 30.63 mmol, 2.00 equiv) dropwise with stirring at room temperature. After the solution was stirred for 1.5 h at room temperature, it was then quenched by the addition of 20 mL of methanol. The resulting mixture was washed with 2×150 mL saturated solution of sodium bicarbonate, and the organic layers concentrated under vacuum. The crude product was purified by Flash-Prep-HPLC with the following conditions (CombiFlash-1): C18 column; mobile phase, 5% ACN/NH₄CO₃ 0.05% in H₂O increasing to 80% ACN/NH₄CO₃ 0.05% in H₂O within 35 min; Detector, UV 254 nm. This resulted in N-[9-[(2R,3R,4R,5R)-5-[[bis(4-methoxyphenyl) (phenyl) methoxy]methyl]-4-([[bis(propan-2-yl)amino](2-cyanoethoxy)phosphanyl]oxy)-3-(trifluoro meth oxy) oxolan-2-yl]-6-oxo-6,9-dihydro-1H-purin-2-yl]-2-methylpropanamide as a white solid. LCMS: (ES, m/z): 924 [M+H]⁺. ¹H-NMR: (300 MHz, DMSO, ppm): δ 12.15 (s, 1H), 11.52 (s, 1H), 8.24 (d, J=1.4 Hz, 1H), 7.42-7.16 (m, 9H), 6.84 (m, 4H), 6.17 (dd, J=18.6, 6.2 Hz, 1H), 5.78 (m, 1H), 4.62-4.45 (m, 1H), 4.26 (m, 1H), 3.74 (d, J=2.0 Hz, 7H), 3.69-3.39 (m, 3H), 3.41 (m, 1H), 3.25 (dd, J=10.7, 3.0 Hz, 1H), 2.83-2.68 (m, 2H), 2.59 (t, J=5.9 Hz, 1H), 1.14 (m, 14H), 1.01 (d, J=6.7 Hz, 4H). ¹⁹FNMR: (300 MHz, DMSO, ppm): δ−57.612, −57.740. ³¹PNMR: (300 MHz, DMSO, ppm):150.775, 150.450.

Step 1: To a slurry of (3R,4R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-3-methyldihydrofuran-2(3H)-one (WO2013/188480, 2013, A1) (50 g, 308 mmol) in DCM (500 mL) was added pyridine (64.8 mL). The mixture was stirred in an ice bath and then benzoyl chloride (95 g, 678 mmol) was added dropwise over 65 minutes at −5° C. The reaction mixture was then warmed to room temperature and stirred for 1.5 h. The reaction mixture was then cooled with an ice bath and water (215 mL) was added. The reaction mixture was warmed to room temperature and stirred for 2 h at this temperature. The organic layer was washed with aqueous hydrogen chloride (1 N, 170 mL), water (170 mL), saturated aqueous sodium carbonate (170 mL), water (3×170 mL) and brine (170 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was treated with toluene (100 mL) and it began to solidify. The solid was filtered and washed with toluene (100 mL) to give ((2R,3R,4R)-3-(benzoyloxy)-4-hydroxy-4-methyl-5-oxotetrahydrofuran-2-yl)methyl benzoate. MS ESI calculated for C20H1807 [M+H]⁺ 371.11, found 371.10. ¹H-NMR: (300 MHz, DMSO-d₆) δ 8.10-7.97 (m, 2H), 7.96-7.84 (m, 2H), 7.72-7.59 (m, 2H), 7.58-7.41 (m, 4H), 6.40 (s, 1H), 5.42 (d, J=7.1 Hz, 1H), 4.98 (td, J=6.4, 3.7 Hz, 1H), 4.67 (dd, J=12.3, 3.7 Hz, 1H), 4.59 (dd, J=12.4, 6.0 Hz, 1H), 1.45 (s, 3H).

Step 2: To a solution of ((2R,3R,4R)-3-(benzoyloxy)-4-hydroxy-4-methyl-5-oxotetrahydrofuran-2-yl)methyl benzoate (49 g, 132 mmol) in DCM (353 mL) was added dropwise diethylaminosulfurtrifluoride (34.1 g, 212 mmol) over 1 h at −78° C. The reaction was slowly warmed to room temperature and stirred for 16 h. The reaction was then quenched with saturated aqueous sodium carbonate (150 mL). The organic layer was washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give ((2R,3R,4S)-3-(benzoyloxy)-4-fluoro-4-methyl-5-oxotetrahydrofuran-2-yl)methyl benzoate. MS ESI calculated for C₂₀H₁₇FO₆ [M+H]⁺ 373.10, found 373.10. ¹H-NMR: (400 MHz, CDCl₃) δ 8.10-8.02 (m, 4H), 7.69-7.63 (m, 1H), 7.63-7.56 (m, 1H), 7.54-7.48 (m, 2H), 7.47-7.41 (m, 2H), 5.87 (dd, J=18.8, 5.3 Hz, 1H), 4.83-4.72 (m, 2H), 4.73-4.62 (m, 1H), 1.79 (d, J=23.2 Hz, 3H). ¹⁹F-NMR: (400 MHz, CDCl₃) δ−153.28 (s, 1F).

Step 3: To a solution of ((2R,3R,4S)-3-(benzoyloxy)-4-fluoro-4-methyl-5-oxotetrahydrofuran-2-yl)methyl benzoate (62 g, 167 mmol) in dry THF (500 mL) was added dropwise lithium tri-tert-butoxyaluminum hydride (1M in THF, 200 mL, 200 mmol) at −78° C. within 30 min. The reaction was warmed to room temperature and stirred for 2 h at this temperature. The mixture was cooled to 0° C. and quenched by saturated aqueous ammonium chloride (800 mL), diluted with ethyl acetate (1000 mL). The resulting mixture was then filtered and the organic layer was washed with brine (800 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give ((2R,3R,4S)-3-(benzoyloxy)-4-fluoro-5-hydroxy-4-methyltetrahydrofuran-2-yl)methyl benzoate which was used directly in next step. MS ESI calculated for C₂₀H₁₉FO₆ [M+H]⁺ 375.12, found 357.10. ¹⁹F-NMR: (282 MHz, CDCl₃) δ−149.67, −167.30.

Step 4: To a solution of ((2R,3R,4S)-3-(benzoyloxy)-4-fluoro-5-hydroxy-4-methyltetrahydrofuran-2-yl)methyl benzoate (67.3 g, 180 mmol) (co-evaporated with dry toluene (100 mL×3) before being used) in dry DCM (565 mL) were added triethylamine (32.5 mL, 234 mmol), N,N-dimethylpyridin-4-amine (4.39 g, 36.0 mmol) and acetic anhydride (25.5 mL, 270 mmol) at 0° C. The reaction mixture was then warmed to room temperature and stirred for 2 h at this temperature. The solvent was removed under reduced pressure and the residue was purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-20%) in petroleum ether to give (2R,3R,4S)-5-acetoxy-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate. MS ESI calculated for C₂₂H₂₁FO₇ [M+NH₄]⁺ 434.13, found 434.20.

Step 5: To a solution of (2R,3R,4S)-5-acetoxy-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (80 g, 192 mmol) in DCM (720 mL) was added hydrogen bromide (33% in acetic acid, 95 mL, 576 mmol) at 0° C. The reaction was stirred at ambient temperature for 16 h. The reaction mixture was quenched by sodium bicarbonate until no carbon dioxide was observed and diluted with DCM (800 mL). The organic layer was then washed with saturated aqueous sodium bicarbonate (3×700 mL), water (700 mL), brine (700 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give ((2R,3R,4S)-3-(benzoyloxy)-5-bromo-4-fluoro-4-methyltetrahydrofuran-2-yl)methyl benzoate which was used directly in next step. MS ESI calculated for C₂₀H₁₈BrFO₅ [M+H]⁺ 437.03, 439.03, found 437.15, 439.05.

Step 6: To a solution of N-(9H-purin-6-yl)benzamide (45.2 g, 189 mmol) in anhydrous dioxane (700 mL) was added sodium hydride (60% in mineral oil, 9.35 g, 234 mmol) at room temperature under argon atmosphere and the mixture was stirred for 5 min at this temperature. Then a solution of ((2R,3R,4S)-3-(benzoyloxy)-5-bromo-4-fluoro-4-methyltetrahydrofuran-2-yl)methyl benzoate (78.6 g, 180 mmol) in anhydrous dioxane (700 mL) was added at room temperature. The reaction mixture was stirred at 100° C. for 5 h. The reaction mixture was quenched by saturated aqueous ammonium chloride (800 mL) and diluted with ethyl acetate (1000 mL). The organic layer was then washed with water (800 mL), brine (800 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a stepwise gradient of 0-60% ethyl acetate in petroleum ether to give a crude product. The crude product was further purified by Combi-Flash with the following conditions: Column: AQ-C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 35 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and evaporated under reduced pressure to give (2R,3R,4S,5R)-5-(6-benzamido-9H-purin-9-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate.

MS ESI calculated for C₃₂H₂₆FN₅O₆ [M+H]⁺ 596.19, found 596.20. ¹H-NMR: (400 MHz, CDCl₃) δ 9.12 (s, 1H), 8.84 (s, 1H), 8.37 (d, J=3.3 Hz, 1H), 8.18-8.09 (m, 4H), 8.09-8.01 (m, 2H), 7.74-7.42 (m, 9H), 6.52 (d, J=20.4 Hz, 1H), 5.68 (dd, J=16.8, 2.6 Hz, 1H), 4.90 (dd, J=12.0, 5.9 Hz, 1H), 4.84 (dd, J=12.0, 4.1 Hz, 1H), 4.60-4.57 (m, 1H), 1.61 (d, J=22.6 Hz, 3H). ¹⁹F-NMR: (376 MHz, CDCl₃) δ−160.17 (s, 1F).

Step 7: To a solution of (2R,3R,4S,5R)-5-(6-benzamido-9H-purin-9-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (10 g, 16.79 mmol) in methanol (70 mL), THF (56 mL) and water (14 mL) was added dropwise aqueous sodium hydroxide solution (4 N, 12.59 mL, 50.4 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 25 min. The reaction mixture was quenched by formic acid (1.89 mL). Then the solvent was removed under reduced pressure. The crude product was purified by column chromatography on silica gel using a stepwise gradient of MeOH (0-10%) in DCM to give N-(9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-9H-purin-6-yl)benzamide. MS ESI calculated for C₁₈H₁₈FN₅O₄ [M+H]⁺ 388.13, found 388.15. ¹H-NMR: (400 MHz, CD₃OD): δ 8.76 (s, 1H), 8.64 (d, J=2.7 Hz, 1H), 8.13-8.06 (m, 2H), 7.71-7.62 (m, 1H), 7.62-7.53 (m, 2H), 6.37 (d, J=16.3 Hz, 1H), 4.33 (dd, J=19.1, 4.5 Hz, 1H), 4.03 (q, J=4.8 Hz, 1H), 3.92 (dd, J=12.1, 4.2 Hz, 1H), 3.87 (dd, J=12.1, 5.5 Hz, 1H), 1.51 (d, J=23.0 Hz, 3H). ¹⁹F-NMR: (376 MHz, CD₃OD): δ−159.16 (s, 1F).

Step 8: To a solution of N-(9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-9H-purin-6-yl)benzamide (8.8 g, 22.72 mmol) in dry pyridine (45 mL) was added 4,4′-(chloro(phenyl)methylene)bis(methoxybenzene) (8.47 g, 24.99 mmol) at room temperature. The reaction mixture was stirred at room temperature for 2 h. The solvent was removed under reduced pressure. The residue was co-evaporated with toluene (3×40 mL) and then purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-100%) in petroleum ether to give a yellow crude product. The crude product was further purified by Combi-Flash with the following conditions: Column: AQ-C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 35 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and roto-evaporated in vacuo to give N-(9-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-9H-purin-6-yl)benzamide. MS ESI calculated for C₃₉H₃₆FN₅O₆ [M+H]⁺ 690.26, found 690.25.

¹H-NMR: (300 MHz, CD₃OD) δ 8.69 (s, 1H), 8.33 (d, J=2.9 Hz, 1H), 8.10-8.01 (m, 2H), 7.65-7.43 (m, 5H), 7.36-7.15 (m, 7H), 6.82 (d, J=8.9 Hz, 4H), 6.33 (d, J=17.0 Hz, 1H), 4.29 (dd, J=18.8, 4.3 Hz, 1H), 4.14-4.10 (m, 1H), 3.74 (s, 6H), 3.46 (dd, J=10.1, 6.4 Hz, 1H), 3.39 (dd, J=10.1, 4.0 Hz, 1H), 1.43 (d, J=23.1 Hz, 3H).

Step 9: To a solution of 3-((bis(diisopropylamino)phosphino)oxy)propanenitrile (8.46 g, 28.1 mmol) (co-evaporated with dry acetonitrile (3×20 mL) before being used) in dry acetonitrile (20 mL) was added pyridin-1-ium 2,2,2-trifluoroacetate (5.42 g, 28.1 mmol) at room temperature. The solution was marked as solution A. To a solution of N-(9-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-9H-purin-6-yl)benzamide (12.9 g, 18.70 mmol) (co-evaporated with dry acetonitrile (3×50 mL) before being used) in dry acetonitrile (45 mL) was added to solution A at room temperature. The reaction was stirred at room temperature for 1 h. The reaction was quenched by water (300 mL) and diluted with ethyl acetate (400 mL). The organic layer was washed with saturated aqueous sodium bicarbonate (300 mL), water (300 mL) and brine (300 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by Combi-Flash with the following conditions: Column: AQ-C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 35 min); Flow rate: 100 m/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and the solvent was removed under reduced pressure to give (2R,3R,4S,5R)-5-(6-benzamido-9H-purin-9-yl)-2-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl (2-cyanoethyl) diisopropylphosphoramidite. MS ESI calculated for C₄₈H₅₃FN₇O₇P [M+H]⁺ 890.37, found 890.45. ¹H-NMR: (400 MHz, DMSO-d₆) δ 11.30 (s, 1H), 8.76 (s, 0.6H), 8.75 (s, 0.4H), 8.44 (d, J=2.3 Hz, 0.4H), 8.40 (d, J=2.4 Hz, 0.6H), 8.11-8.04 (m, 2H), 7.71-7.62 (m, 1H), 7.61-7.53 (m, 2H), 7.52-7.40 (m, 2H), 7.35-7.20 (m, 7H), 6.90-6.85 (m, 4H), 6.39 (d, J=16.0 Hz, 0.4H), 6.38 (d, J=16.0 Hz, 0.6H), 4.76-4.61 (m, 1H), 4.31-4.19 (m, 1H), 3.82-3.70 (m, 1H), 3.74 (s, 6H), 3.66-3.43 (m, 5H), 2.75 (t, J=5.8 Hz, 1H), 2.59 (t, J=5.9 Hz, 1H), 1.59 (d, J=22.9 Hz, 1.5H), 1.53 (d, J=23.0 Hz, 1.5H), 1.23-0.96 (m, 12H). ¹⁹F-NMR: (376 MHz, DMSO-d₆) δ−154.29 (d, J=3.7 Hz, 0.5F), −154.91 (s, 0.5F). ³¹P-NMR: (162 MHz, DMSO-d₆) δ 150.40 (d, J=3.7 Hz, 0.5P), 150.14 (s, 0.5P).

Step 1: To a suspension of N-(2-oxo-1,2-dihydropyrimidin-4-yl)benzamide (88 g, 408 mmol) in dry acetonitrile (200 mL) was added (E)-trimethylsilyl N-(trimethylsilyl)acetimidate (225 mL, 919 mmol) dropwise at room temperature under an argon atmosphere. The mixture was then heated to 70° C. and stirred for 2 h. The reaction was cooled to room temperature and a solution of (2R,3R,4S)-5-acetoxy-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (85 g, 204 mmol) (co-evaporated with dry acetonitrile (3×50 mL) before being used) in dry acetonitrile (225 mL) was added at room temperature. The mixture was heated to 70° C. and stirred for 10 h. The reaction was diluted with ethyl acetate (1500 mL) and quenched by saturated aqueous sodium bicarbonate (600 mL) at 0° C. and the mixture was filtered. The organic layer was washed with saturated sodium bicarbonate (3×600 mL), water (600 mL) and brine (800 mL). The organic layer was dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-50%) in petroleum ether to give (2R,3R,4S,5R)-5-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate. MS ESI calculated for C₃₁H₂₆FN₃O₇ [M+H]⁺ 572.18, found 572.15. ¹H-NMR: (400 MHz, DMSO-d₆) δ 11.36 (s, 1H), 8.18-7.99 (m, 7H), 7.81-7.48 (m, 9H), 7.41 (d, J=7.6 Hz, 1H), 6.42 (d, J=17.6 Hz, 1H), 5.60 (dd, J=19.7, 4.6 Hz, 1H), 4.80-4.74 (m, 2H), 4.68-4.61 (m, 1H), 1.60 (d, J=23.5 Hz, 3H). ¹⁹F-NMR: (376 MHz, DMSO) δ−158.48 (s, 1F).

Step 2: To a solution of (2R,3R,4S,5R)-5-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (14.1 g, 24.67 mmol) in THF (99 mL), MeOH (79 mL) and water (19.7 mL) was added dropwise aqueous sodium hydroxide (4 N, 18.50 mL, 74.0 mmol) at 0° C. The reaction was then stirred at room temperature for 20 min. The reaction was quenched by formic acid (2.82 mL, 74.0 mmol) at 0° C. The solvent was then removed under reduced pressure and the residue was purified by column chromatography on silica gel using a stepwise gradient of MeOH (0-10%) in DCM to give N-(1-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)benzamide. MS ESI calculated for C17H₁₈FN₃O₅ [M+H]⁺ 364.12, found 364.15. ¹H-NMR: (400 MHz, DMSO-d₆) δ 11.31 (s, 1H), 8.23 (d, J=7.6 Hz, 1H), 8.06-7.97 (m, 2H), 7.69-7.60 (m, 1H), 7.62-7.49 (m, 2H), 7.41-7.36 (m, 1H), 6.09 (d, J=13.8 Hz, 1H), 5.93 (d, J=5.7 Hz, 1H), 5.12 (t, J=5.6 Hz, 1H), 4.13-4.05 (m, 1H), 3.82-3.78 (m, 1H), 3.77-3.72 (m, 1H), 3.68-3.62 (m, 1H), 1.46 (d, J=23.6 Hz, 3H). ¹⁹F-NMR: (376 MHz, DMSO-d₆) δ−156.84 (s, 1F).

Step 3: To a solution of N-(1-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)benzamide (7.2 g, 19.82 mmol) (co-evaporated with dry pyridine (3×20 mL) before being used) in dry pyridine (36 mL) was added 4,4′-(chloro(phenyl)methylene)bis(methoxybenzene) (7.39 g, 21.80 mmol) at room temperature. The reaction was stirred at room temperature for 2 h. The solvent was removed under reduced pressure and the residue was co-evaporated with toluene (2×15 mL). The residue was purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-100%) in petroleum ether to give a crude product. The crude product was further purified by Combi-Flash with the following conditions: Column: AQ C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 35 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and the solvent was removed under reduced pressure to give N-(1-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)benzamide. MS ESI calculated for C₃₈H₃₆FN₃O₇ [M+H]⁺ 666.25, found 666.25. ¹H-NMR: (400 MHz, DMSO-d₆) δ 11.32 (s, 1H), 8.05-8.01 (m, 3H), 7.69-7.60 (m, 1H), 7.58-7.49 (m, 2H), 7.44 (d, J=7.8 Hz, 2H), 7.38-7.26 (m, 8H), 6.97-6.90 (m, 4H), 6.13 (d, J=13.2 Hz, 1H), 6.03 (d, J=5.7 Hz, 1H), 4.16 (dt, J=21.4, 6.0 Hz, 1H), 4.01-3.97 (m, 1H), 3.77 (s, 6H), 3.42-3.33 (m, 2H), 1.46 (d, J=23.6 Hz, 3H). ¹⁹F-NMR: (376 MHz, DMSO-d₆) δ−156.48 (s, 1F).

Step 4: To a solution of 3-(((diisopropylamino)(2,4-dimethylpentan-3-yl)phosphino)oxy)propanenitrile (6.50 g, 21.63 mmol) (co-evaporated with dry acetonitrile (3×20 mL) before being used) in dry acetonitrile (20 mL) was added pyridine 2,2,2-trifluoroacetate (4.18 g, 21.63 mmol) at room temperature. After 5 min, a solution of N-(1-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)benzamide (9.6 g, 14.42 mmol) (co-evaporated with dry acetonitrile (3×20 mL) before being used) in dry acetonitrile (30 mL) was added at room temperature. The reaction was stirred at room temperature for 1.5 h. The reaction was diluted with ethyl acetate (500 mL) and quenched by saturated aqueous sodium bicarbonate (300 mL). The organic layer was then washed with water (300 mL) and brine (300 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was further purified by Combi-Flash with the following conditions: Column: AQ C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 35 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and roto-evaporated in vacuo to give (2R,3R,4S,5R)-5-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-2-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl (2-cyanoethyl) diisopropylphosphoramidite. MS ESI calculated for C₄₇H₅₃FN₅O₈P [M+H]⁺ 866.36, found 866.45. ¹H-NMR: (400 MHz, DMSO-d₆) δ 11.35 (s, 1H), 8.12 (d, J=7.9 Hz, 0.5H), 8.10 (d, J=8.0 Hz, 0.5H), 8.05-7.98 (m, 2H), 7.69-7.60 (m, 1H), 7.58-7.49 (m, 2H), 7.49-7.39 (m, 2H), 7.38-7.26 (m, 8H), 6.97-6.88 (m, 4H), 6.23-6.12 (m, 1H), 4.51-4.41 (m, 1H), 4.16-4.11 (m, 1H), 3.77 (s, 6H), 3.59-3.40 (m, 6H), 2.75 (t, J=5.9 Hz, 1H), 2.59 (t, J=6.0 Hz, 1H), 1.58 (d, J=14.1 Hz, 1.4H), 1.52 (d, J=14.0 Hz, 1.6H), 1.22-0.93 (m, 12H). ¹⁹F-NMR: (376 MHz, DMSO-d₆) δ−155.59 (s, 0.5F), −155.99 (s, 0.5F). ³¹P-NMR: (162 MHz, DMSO-d₆) δ 150.64 (s, 0.5P), 150.46 (s, 0.5P).

Step 1: To a solution of uracil (16.9 g, 144 mmol) in acetonitrile (150 mL) was added N,O-bis(trimethylsilyl)acetamide (84 mL, 324 mmol) at 85° C. The solution was stirred at 85° C. for 2 h and then cooled to room temperature. A solution of (2R,3R,4S)-5-acetoxy-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (30.0 g, 72 mmol) in acetonitrile (150 mL) was added followed by dropwise addition of trimethylsilyl trifluoromethanesulfonate (84 g, 360 mmol) at room temperature. The reaction mixture was slowly warmed to 85° C. and stirred for 16 h. The mixture was then cooled to 0° C., quenched with saturated sodium bicarbonate solution (500 mL). The mixture was diluted with ethyl acetate (500 mL). The separated organic layer was washed with brine (3×100 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with 10%-50% ethyl acetate in petroleum ether to afford ((2R,3R,4S)-3-(benzoyloxy)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-4-methyltetrahydrofuran-2-yl)methyl benzoate. MS ESI calculated for C₂₄H₂₁FN₂O₇ [M+H]⁺ 469.13, found 469.15. ¹H-NMR (300 MHz, CDCl₃) δ 9.48 (brs, 0.5H), 9.40 (brs, 0.5H), 8.21-7.87 (m, 5H), 7.76-7.36 (m, 6H), 6.56 (d, J=15.5 Hz, 0.5H), 6.30 (d, J=20.3 Hz, 0.5H), 5.86-5.70 (m, 1H), 5.62-5.45 (m, 1H), 5.00-4.57 (m, 3H), 1.66 (d, J=11.4, 1.5H), 1.56 (d, J=11.4 Hz, 1.5H).

Step 2: To a solution of ((2R,3R,4S)-3-(benzoyloxy)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-4-methyltetrahydrofuran-2-yl)methyl benzoate (48 g, 103 mmol) in MeOH (125 mL), THF (100 mL) and H₂O (25 mL) was added aqueous sodium hydroxide solution (77.8 mL, 4 M, 309 mmol) at 0° C. The mixture was stirred at room temperature for 30 min. The pH of the mixture was adjusted to 7 with formic acid and the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with 0%-20% MeOH in DCM to afford 1-((3S, 4R, 5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H, 3H)-dione as a white solid. The product (containing α and β isomers) was separated by Prep-SFC with the following conditions: (Column: ChiralPak IA-SFC, 5 cm×25 cm, 5 um; Mobile Phase A: CO₂: 50, Mobile Phase B: MeOH: 50; Flow rate:180 mL/min; 220 nm; RT1: 2.51 min (β isomer); RT2: 3.46 min (a isomer)) to give 1-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione. MS ESI calculated for C₁₀H₁₃FN₂O₅ [M+H]⁺ 261.08, found 261.10. ¹H-NMR (300 MHz, CD₃OD) δ 7.82 (dd, J=8.2, 2.4 Hz, 1H), 6.03 (d, J=15.9 Hz, 1H), 5.70 (d, J=8.2 Hz, 1H), 4.13 (dd, J=20.7, 4.7 Hz, 1H), 3.92-3.79 (m, 2H), 3.84-3.68 (m, 1H), 1.48 (d, J=23.3 Hz, 3H).

Step 3: To a solution of 1-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione (9.5 g, 365.0 mmol) in pyridine (60 mL) was added 4,4′-(chloro(phenyl)methylene)bis(methoxybenzene) (13.6 g, 402.0 mmol) at 0° C. The mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. The residue was purified by Combi Flash Column Chromatography as the following conditions: (Column: AQ-C¹⁸, 19×250 mm, 10 um; Mobile Phase A: Water, Mobile Phase B: MeCN; Flow rate: 100 mL/min; Gradient: 30% B hold 5 min, 30% B to 100% B in 35 min; 100% B hold 8 min, 254/280 nm). The product-containing fractions were pooled and concentrated under reduced pressure to afford 1-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione. MS ESI calculated for C₃₁H₃₁FN₂O₇ [M−H]⁻ 561.21, found 561.21. ¹H-NMR (400 MHz, CD₃OD) δ 7.78-7.70 (m, 1H), 7.50-7.43 (m, 2H), 7.39-7.19 (m, 7H), 6.93-6.82 (m, 4H), 6.03 (d, J=14.1 Hz, 1H), 5.47 (dd, J=8.0, 2.1 Hz, 1H), 4.27 (dd, J=20.7, 5.6 Hz, 1H), 3.96 (q, J=4.7 Hz, 1H), 3.83-3.74 (m, 6H), 3.44 (d, J=4.2 Hz, 2H), 1.48 (d, J=23.3 Hz, 3H).

Step 4: 1-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione (15.0 g, 26.6 mmol) was co-evaporated with dry acetonitrile (3×10 mL) and then re-dissolved in DCM (40 mL), marked as solution A which was kept under an argon atmosphere before being used. 3-(Bis(diisopropylamino)phosphinooxy)propanenitrile (12.0 g, 39.9 mmol) was also co-evaporated with dry acetonitrile (3×10 mL) and then re-dissolved in DCM (35 mL), marked as solution B. 4,5-Dicyanoimidazole (2.6 g, 22.0 mmol) was added into the solution B, followed by the addition of solution A at ambient temperature. The resulting mixture was stirred at ambient temperature for 2 h. The reaction mixture was diluted with DCM (300 mL) and washed with saturated aqueous sodium bicarbonate solution (300 mL) and the organic layer was concentrated under reduced pressure. The residue was purified by Combi Flash Column Chromatography as the following conditions: (Column: AQ-C¹⁸, 19×250 mm, 10 um; Mobile Phase A: Water, Mobile Phase B: MeCN; Flow rate: 100 mL/min; Gradient: 30% B hold 5 min, 30% B to 100% B in 30 min; 100% B hold 8 min, 254/280 nm). The product-containing fractions were pooled and concentrated under reduced pressure to afford (2R,3R,4S,5R)-2-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-fluoro-4-methyltetrahydrofuran-3-yl (2-cyanoethyl) diisopropylphosphoramidite. MS ESI calculated for C₄₀H₄₈FN₄O₈P [M+H]⁺ 763.32, found 763.30. ¹H-NMR (300 MHz, DMSO-d₆) δ 11.55 (s, 1H), 7.63-7.60 (m, 1H), 7.46-7.42 (m, 2H), 7.37-7.24 (m, 7H), 6.94-6.85 (m, 4H), 6.05-5.95 (m, 1H), 5.60-5.46 (m, 1H), 4.47-4.33 (m, 1H), 4.15-4.03 (m, 1H), 3.85-3.65 (m, 1H), 3.76 (s, 6H), 3.65-3.29 (m, 5H), 2.75 (t, J=5.8 Hz, 1H), 2.58 (t, J=5.9 Hz, 1H), 1.55-1.43 (m, 3H), 1.14-0.98 (m, 12H). ¹⁹F-NMR (282 MHz, DMSO-d₆): δ−155.85 (s, 0.3F), −156.32 (s, 0.7F). ³¹P-NMR (121 MHz, DMSO-d₆): δ 150.57 (s, 0.4P), 150.23 (s, 0.6P).

Step 1: To a suspension of ((2R,3R,4S)-3-(benzoyloxy)-4-fluoro-5-hydroxy-4-methyltetrahydrofuran-2-yl)methyl benzoate (74 g, 198 mmol), 6-chloro-9H-purin-2-amine (36.9 g, 217 mmol) and triphenylphosphine (156 g, 593 mmol) in dry THF (740 mL) was added (E)-diethyl diazene-1,2-dicarboxylate (93 mL, 593 mmol) at 0° C. The reaction was then stirred at ambient temperature for 3 h. After completion of the reaction, the solvent was removed and the residue was diluted with ethyl acetate (1000 mL). The organic layer was then washed with saturated aqueous sodium bicarbonate solution (2×500 mL), water (500 mL) and brine (500 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was then purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-50%) in petroleum ether to give product which is a mixture of α and β isomers. The crude product was used as it. MS ESI calculated for C₄₃H₃₄ClFN₅O₅P [M+H]⁺ 786.20, 788.20, found 786.25, 788.25.

Step 2: To a suspension of ((2R,3R,4S,5R)-3-(benzoyloxy)-5-(6-chloro-2-((triphenylphosphoranylidene)amino)-9H-purin-9-yl)-4-fluoro-4-methyltetrahydrofuran-2-yl)methyl benzoate (133 g, 169 mmol) in EtOH (700 mL) was added acetic acid (700 mL, 1 M in water, 700 mmol) at ambient temperature. The reaction was then stirred at 80° C. for 2 h. After completion of the reaction, the solvent was removed and the residue was diluted with ethyl acetate (1000 mL) and neutralized with sodium bicarbonate (200 g). The organic layer was then washed with saturated aqueous sodium bicarbonate solution (6×500 mL), water (500 mL) and brine (500 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-50%) in petroleum ether to give a crude product. To the crude product was then added Et₂O (300 mL). The resulting solid, which contains triphenylphosphine oxide and most of the a isomer, was filtered off and the filtrate was concentrated under reduced pressure. The residue was then purified by column chromatography on silica gel using a stepwise gradient of ethyl acetate (0-50%) in petroleum ether to give (2R,3R,4S,5R)-5-(2-amino-6-chloro-9H-purin-9-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate. MS ESI calculated for C₂₅H₂₁ClFN₅O₅ [M+H]⁺ 526.12, 528.12, found 526.20, 528.20. ¹H-NMR: (300 MHz, DMSO-d₆) δ 8.14 (d, J=3.2 Hz, 1H), 8.13 (m, 4H), 7.79-7.51 (m, 6H), 7.12 (brs, 2H), 6.25 (d, J=18.8 Hz, 1H), 5.71 (dd, J=18.9, 4.0 Hz, 1H), 4.83-4.69 (m, 2H), 4.69-4.65 (m, 1H), 1.55 (d, J=23.1 Hz, 3H). ¹⁹F-NMR: (282 MHz, DMSO-d₆) δ−157.00 (d, J=1.9 Hz, 1F).

Step 3: To (2R,3R,4S,5R)-5-(2-amino-6-chloro-9H-purin-9-yl)-2-((benzoyloxy)methyl)-4-fluoro-4-methyltetrahydrofuran-3-yl benzoate (22 g, 41.8 mmol) was added ammonia (200 mL, 7 M in 2-propanol, 1400 mmol) at −78° C. The reaction was then stirred at 90° C. for 2 d. After completion of the reaction, the solvent was removed and ethyl acetate (150 mL) was added. The suspension was filtered and the filter cake was washed with ethyl acetate (30 mL) to give (2R,3R,4S,5R)-5-(2,6-diamino-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol. MS ESI calculated for C₁₁H₁₅FN₆O₃ [M+H]⁺ 299.12, found 299.10.

¹H-NMR: (300 MHz, DMSO-d₆): δ 7.77 (d, J=3.1 Hz, 1H), 6.80 (brs, 2H), 6.02 (s, 1H), 5.88 (brs, 2H), 5.86 (d, J=17.9 Hz, 1H), 5.07 (s, 1H), 4.15 (d, J=19.3 Hz, 1H), 3.84-3.73 (m, 1H), 3.73-3.55 (m, 2H), 1.36 (d, J=23.1 Hz, 3H). ¹⁹F-NMR: (282 MHz, DMSO-d₆): δ−157.21 (s, 1F).

Step 4: To a solution of (2R,3R,4S,5R)-5-(2,6-diamino-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol (11.5 g, 38.6 mmol) in Tris-buffer (193 mL, pH=7.5) was added adenosine deaminase (193 mg, 38.6 mmol) at ambient temperature. The mixture was then warmed to 40° C. and stirred for 16 h. The mixture was concentrated. The residue was purified by Combi-Flash Column Chromatography with the following conditions: Column: AQ C18gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 0-20% within 50 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and the solvent was removed under reduced pressure to give 2-amino-9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-1H-purin-6(9H)-one. MS ESI calculated for C₁₁H₁₄FN₅O₄ [M+H]⁺ 300.10, found 300.10. ¹H-NMR: (300 MHz, DMSO-d₆): δ 7.58 (d, J=3.1 Hz, 1H), 7.01 (brs, 2H), 6.16 (brs, 1H), 5.77 (d, J=17.6 Hz, 1H), 5.16 (brs, 1H), 4.11 (dd, J=19.3, 4.5 Hz, 1H), 3.79-3.66 (m, 1H), 3.62-3.54 (m, 2H), 1.31 (d, J=23.0 Hz, 3H). ¹⁹F-NMR: (282 MHz, DMSO-d₆): δ−157.34 (s, 1F).

Step 5: To a suspension of 2-amino-9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-1H-purin-6(9H)-one (6.2 g, 20.72 mmol) in dry pyridine (31 mL) was added trimethylsilyl chloride (9.6 mL, 103.7 mmol) at 0° C. The reaction was then warmed to room temperature and continued to stir for 2 h. Then isobutryic anhydride (3.2 mL, 31.1 mmol) was added at 0° C. The mixture was stirred at room temperature for 1.5 h. After completion of the reaction, ammonium hydroxide (100 mL) was added. The solvents were removed under reduced pressure and the residue was purified by chromatography on silica gel using a stepwise gradient of MeOH (0-20%) in DCM to give N-(9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-2-yl)isobutyramide. MS ESI calculated for C₁₅H₂₀FN₅O₅ [M+H]⁺ 370.14, found 370.10.

¹H-NMR: (400 MHz, DMSO-d₆) δ 12.13 (s, 1H), 11.67 (s, 1H), 8.09 (d, J=2.7 Hz, 1H), 6.06 (d, J=5.1 Hz, 1H), 5.91 (d, J=15.8 Hz, 1H), 5.10 (t, J=5.7 Hz, 1H), 4.20 (dt, J=18.5, 4.9 Hz, 1H), 3.88-3.79 (m, 1H), 3.76-3.57 (m, 2H), 2.78 (h, J=6.8 Hz, 1H), 1.40 (d, J=23.2 Hz, 3H), 1.13 (d, J=6.8 Hz, 6H). ¹⁹F-NMR: (282 MHz, DMSO-d₆): δ−157.74 (s, 1F).

Step 6: To a solution of N-(9-((2R,3S,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-2-yl)isobutyramide (3 g, 8.12 mmol) in dry pyridine (15 mL) was added 4,4′-(chloro(phenyl)methylene)bis(methoxybenzene) (3.02 g, 8.9 mmol) at room temperature. The reaction was quenched by adding MeOH (5 mL). The solvent was removed and the residue was purified by Combi-Flash Colum Chromatography with the following conditions: Column: AQ C18gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 60-100% within 50 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and the solvent was removed under reduced pressure to give N-(9-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-2-yl)isobutyramide. MS ESI calculated for C₃₆H₃₈FN₅O₇ [M+H]⁺ 672.28, found 672.25. ¹H-NMR: (400 MHz, DMSO-d₆) δ 12.15 (s, 1H), 11.68 (s, 1H), 7.86 (d, J=2.9 Hz, 1H), 7.48-7.38 (m, 2H), 7.35-7.18 (m, 7H), 6.93-6.81 (m, 4H), 6.09 (d, J=5.0 Hz, 1H), 5.97 (d, J=16.8 Hz, 1H), 4.20 (dt, J=18.8, 4.8 Hz, 1H), 4.08-3.98 (m, 1H), 3.74 (s, 6H), 3.44-3.29 (m, 1H), 3.23 (dd, J=10.3, 3.6 Hz, 1H), 2.78 (septet, J=6.8 Hz, 1H), 1.39 (d, J=23.2 Hz, 3H), 1.13 (dd, J=6.8, 1.7 Hz, 6H). ¹⁹F-NMR: (376 MHz, DMSO-d₆): δ−157.52 (s, 1F).

Step 7: To a solution of 3-((bis(diisopropylamino)phosphino)oxy)propanenitrile (11.30 g, 37.5 mmol)(co-evaporated with dry MeCN (3×60 mL) before being used) in dry DCM (24 mL) was added 4,5-dicyanoimidazole (4.4 g, 37.5 mmol) at room temperature. The solution was marked as solution A. To a solution of N-(9-((2R,3S,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-3-fluoro-4-hydroxy-3-methyltetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-2-yl)isobutyramide (16.8 g, 25.01 mmol) (co-evaporated with dry MeCN (3×60 mL) before being used) in dry DCM (60 mL) was added solution A at room temperature under argon. The reaction was then stirred at room temperature for 1 h. After completion of the reaction, the reaction mixture was diluted with DCM (300 mL). The organic layer was then washed with saturated aqueous sodium bicarbonate solution (150 mL), water (150 mL) and brine (150 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was then purified by Combi-Flash Column Chromatography with the following conditions: Column: AQ C18 gel column (330 g), 20-40 μm; Mobile Phase: MeCN and water (Gradient: 45-100% within 45 min); Flow rate: 100 mL/min; Detector: UV 254 & 280 nm. The product-containing fractions were collected and roto-evaporated in vacuo to give (2R,3R,4S,5R)-2-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-fluoro-5-(2-isobutyramido-6-oxo-1H-purin-9(6H)-yl)-4-methyltetrahydrofuran-3-yl (2-cyanoethyl) diisopropylphosphoramidite. MS ESI calculated for C₄₅H₅₅FN₇O₈P [M+H]⁺ 872.38, found 872.40. ¹H-NMR: (400 MHz, DMSO-d₆) δ 12.19 (s, 1H), 11.69 (s, 0.4H), 11.66 (s, 0.6H), 7.97-7.90 (m, 1H), 7.48-7.39 (m, 2H), 7.37-7.20 (m, 7H), 6.96-6.85 (m, 4H), 6.04-5.94 (m, 1H), 4.58-4.38 (m, 1H), 4.27-4.12 (m, 1H), 3.86-3.66 (m, 7H), 3.66-3.46 (m, 3H), 3.45-3.27 (m, 2H), 2.87-2.72 (m, 2H), 2.64-2.55 (m, 1H), 1.58-1.41 (m, 3H), 1.18-1.09 (m, 16H), 1.00 (d, J=6.7 Hz, 2H). ¹⁹F-NMR: (376 MHz, DMSO-d₆): δ−155.28 (s, 0.4F), −155.50 (s, 0.6F). ³¹P-NMR: (162 MHz, DMSO-d₆) δ 150.98 (s, 0.4P), 150.03 (s, 0.6P).

TABLE 2 Exemplary nomenclature for first position nucleotides starting at the 5′ end include:

  rG

  rGs

Wherein L is selected from the Table 2a:

Also included within Table 2 above are first position nucleotides where “r” in “rG” or “rGs” represent groups comprising “ome”, “flu,” “thior,” “d,” “fara,” “1na,” “moe,” “4alkd,” “ara,” “meF” “cf3,” 25, rxylo, ami, ome4S, fara8az, and “me,” as illustrated herein.

TABLE 2a L Definition 5′ end substituents L Structure HO

pp

pcp

ppp

p

An embodiment of this invention is realized when the first position nucleotide starting at the 5′ end of Formula Ia and Formula I is referred to as rG wherein V, Y, Z, W, and X are O, —OH, —CH—, N, and O, respectively. An embodiment of this invention is realized when the first position nucleotide starting at the 5′ end of Formula Ia and Formula I is referred to as rGs wherein V, Y, Z, W, and X are O, —SH, —CH—, N, and O, respectively.

Table 3: Nomenclature for internally-located nucleotides (i.e., positions 2-23) contained within the single-stranded molecules exemplified in Table 5. For the purposes of this invention, in Table 3 the terms “r,” “ome”, “flu,” “thior,” “d,” “fara,” “1na,” “moe,” “4alkd,” “ara,” “meF” “cf3,” 25, rxylo, ami, ome4S, fara8az, or “me,” appearing before the “B” in the first and third column of Table 3 represents a sugar group as exemplified in columns two and four of Table 3, and “B” represents a nucleobase or derivative thereof selected from adenine, cytosine, guanine and uracil, which are designated “A”, “C”, “G”, and “U”, respectively.

TABLE 3 Internal nucleotide nomenclature (positions 2-23) Structure rB B = nucleobase A, C, G, U, or I where “I” in “rI” is an “A” analog

omeB B = nucleobase A, C, G, U

fluB B = nucleobase A, C, G, U

thiorB B = nucleobase A, C, G, U

dB B = nucleobase A, C, G, U

faraB B = nucleobase A, C, G, U

lnaB B = nucleobase A, 5-Me-C, G,5- Me-U

moeB B = nucleobase A, 5-Me-C, G,5- Me-U

4alkdB B = nucleobase A, C, G, U

araB B = nucleobase A, C, G, U

meFB B = nucleobase A, C, G, U

cf3B B = nucleobase A, C, G, U

25B B = nucleobase A, C, G, U

rxyloB B = nucleobase A, C, G, U

amiB B = nucleobase A, C, G, U

ome4SB B = nucleobase A, C, G, U

rBs B = nucleobase A, C, G, U, or I where “I” in “rI” is an “A” analog

omeBs B = nucleobase A, C, G, U

fluBs B = nucleobase A, C, G, U

thiorBs B = nucleobase A, C, G, U

dBs B = nucleobase A, C, G, U

faraBs B = nucleobase A, C, G, U

lnaBs B = nucleobase A, 5-Me-C, G,5- Me-U

moeBs B = nucleobase A, 5-Me-C, G,5- Me-U

4alkdBs B = nucleobase A, C, G, U

araBs B = nucleobase A, C, G, U

meFBs B = nucleobase A, C, G, U

cf3Bs B = nucleobase A, C, G, U

25Bs B = nucleobase A, C, G, U

rxyloBs B = nucleobase A, C, G, U

amiBs B = nucleobase A, C, G, U

ome4SBs B = nucleobase A, C, G, U

!TABLE 3a Position 2-23 nucleotides or derivatives thereof Nucleotide fara8azA

fara7dzA

r2AP

d2aminoA

r7dz8azA

4S8azG

d5propU

d5propC

r7dzA

d7dzA

fara8azAs

fara7dzAs

r2APs

d2aminoAs

r7dz8azAs

4S8azGs

d5propUs

d5propCs

r7dzAs

d7dzAs

An embodiment of this invention is realized when nucleotide position 2 starting from the 5′ end is selected from the group consisting of rC, rCs, fluC, fluCs, OMeC, and OMeCs. A subembodiment of this aspect of the invention is realized when nucleotide position 2 starting from the 5′ end is rC or rCs. Another subembodiment of this aspect of the invention is realized when nucleotide position 2 starting from the 5′ end is fluC or fluCs. Another subembodiment of this aspect of the invention is realized when nucleotide position 2 starting from the 5′ end is OMeC or OMeCs.

TABLE 4 Nomenclature for nucleotide position 24 starting from 5′ exemplified in Table 5 Nucleotide Nucleotide position 24 - Structure Position 24 Structure rCSup

rC;pSup

fluC Sup

fluC;pSup

omeCSup

omeC;pSup

An embodiment of this invention is realized when R₃ of the position 24 nucleotide is hydrogen, K is hydrogen or P(O)(OH)₂, X is O, and one of R₁ and R₂ is hydrogen while the other is selected from F, OH, and OMe. An embodiment of this invention is realized when the position 24 nucleotide is selected from Table 4. A subembodiment of this aspect of the invention is realized when nucleotide position 24 is rCSup or rC;pSup. Another subembodiment of this aspect of the invention is realized when nucleotide position 24 starting from the 5′ end is fluCSup or fluC;pSup. Another subembodiment of this aspect of the invention is realized when nucleotide position 24 starting from the 5′ end is omeCSup or omeC;pSup.

An embodiment of this invention is realized when position 23 nucleotide starting from the 5′ end of Formula Ia and Formula I is selected from rG, rGs, fluG, fluGs, omeG, and omeGs. A subembodiment of this aspect of the invention is realized when nucleotide position 23 is rG or rGs. Another subembodiment of this aspect of the invention is realized when nucleotide position 23 starting is fluG or fluGs. Another subembodiment of this aspect of the invention is realized when nucleotide position 23 is omeG or omeGs.

Another embodiment of this invention is realized when starting from the 5′ end of Formula Ia and Formula I, position 1 nucleotide is rG or rGs, and position 24 nucleotide is selected from the group consisting of rCSup, rC;pSup, fluCSup, fluC;pSup, omeC;pSup, and omeCSup.

Another embodiment of this invention is realized when starting from the 5′ end of Formula Ia and Formula I, position 1 nucleotide is rG or rGs, position 2 nucleotide is selected from the group consisting of rC, rCs, fluC, fluCs, omeCs, and omeC, position 23 is selected from the group consisting of rG, rGs, fluG, fluGs, omeG, and omeGs, and position 24 nucleotide is selected from the group consisting of rCSup, rC;pSup, fluCSup, fluC;pSup, omeC;pSup, and omeCSup.

General Synthetic Scheme

Oligonucleotides were prepared by chemical synthesis, as presented in Scheme 1. The oligonucleotide target sequence was synthesized on solid support, such as on controlled pore glass. After cleavage of the protected oligonucleotide from the solid support and deprotection of all protecting groups, each strand was purified chromatographically with a reversed phase (C18) or anion exchange (SAX) resin. After purification, each oligonucleotide product was lyophilized to dryness.

The synthesis of each oligonucleotide was accomplished on a solid support, such as controlled pore glass, using a commercially available automated oligosynthesizers. The solid support was obtained pre-loaded with the first (3′) nucleoside unit of the desired sequence and placed in an appropriate column position for the oligosynthesizer. The first nucleoside was linked to the solid support via a succinate linkage and contained a suitable acid sensitive protecting group (e.g., trityl, dimethoxytrityl) on the 5′-hydroxyl group. The solid-phase oligosynthesis employed synthetic procedures that are generally known in the art. Elongation of the desired oligomeric sequence went through a cycle of four steps: 1) Acidic deprotection of the 5′-trityl protecting group; 2) Coupling of the next nucleotide unit as the 5′-trityl (or dimethoxytrityl) protected phosphoramidite in the presence of an activating agent, such as S-ethyl-tetrazole; 3) Oxidation of the P(III) phosphite triester to the P(V) phosphate triester by an oxidizing agent, such as iodine; and 4) Capping any remaining unreacted alcohol groups through esterification with an acylating agent, such as acetic anhydride. The phosphoramidites used were either derived from naturally occurring nucleotide units or from chemically modified versions of these nucleotides. Typically, all phosphoramidites were prepared in solutions of acetonitrile (or other suitable solvents or solvent mixtures, such as acetonitrile with some percentage of dimethyl formamide or some other appropriate organic solvent). The activator for phosphoramidite coupling was typically dissolved in acetonitrile. An oxidizing agent, such as iodine, was dissolved in a suitable solvent mixture, such as acetonitrile, pyridine and water. Acidic detritylation reagents, such as dichloroacetic acid or trichloroacetic acid, were dissolved in appropriate solvents, such as toluene or dichloromethane. Acylating capping reagents were, for example, a mixture of acetic anhydride, 2,6-lutidine and N-methyl-imidazole in acetonitrile. In place of the oxidation of the phosphite triester to the phosphate triester, the P(III) intermediate may be converted to the phosphorothioate analog with a sulfurizing reagent, such as phenyl-acetyl-disulfide, in a suitable solvent, such as a mixture of acetonitrile and pyridine. In between each step of the oligonucleotide elongation, acetonitrile (or another suitable solvent) was used to remove excess reagents and wash the solid support.

Oligonucleotide synthesis cycles were continued until the last (5′) nucleotide unit was installed onto the extended oligomer. After the final cycle, the 5′-trityl protecting group may or may not be removed from the oligonucleotide while it remains on the solid support. In some instances, the 5′-terminal trityl group was first removed by treatment with an acidic solution. After this deprotection, the solid support was treated with an appropriate base, such as aqueous methyl amine or ammonia:methyl amine (1:1) (at either room temperature or with mild heating) in order to cleave the oligonucleotide from the support, remove the cyanoethyl protecting groups on the phosphate linkages and deprotect the acyl protecting groups on the nucleotide bases. Purification of these oligonucleotides would then be accomplished by SAX or reversed phase (C18 or C8) chromatography. Typically, the oligonucleotide was eluted from the SAX resin with a gradient of an inorganic salt, such as sodium chloride or sodium perchlorate. Salt was removed from the purified samples by dialysis or tangential flow filtration. The desalted material was then lyophilized or annealed. Alternatively, the purified oligonucleotides were annealed prior to removal of salt and then dialyzed.

In order to synthesize certain 5′-modified oligonucleotides, such as those containing 5′-ppp and 5′-pp, additional synthetic steps were performed on the oligonucleotide prior to cleavage from the solid support. After removal of the 5′-protecting group of the terminal nucleotide, a suitable phosphoramidite or comparable reagent was used to generate an activated monophosphate or phosphite intermediate. The intermediate was oxidized, followed by coupling with pyrophosphate or phosphate, or analogs thereof, to provide the desired 5′-ppp or 5′-pp oligonucleotide after cleavage and deprotection of the oligonucleotide under appropriate conditions. Alternatively, the intermediate was coupled to pyrophosphate or phosphate, or analogs thereof, followed by the oxidation step.

In other instances, the 5′-trityl protecting group was left on the oligonucleotide during basic cleavage from the solid support and deprotection of the oligonucleotide protecting groups. In this case, the oligonucleotide was purified with a reversed phase resin, such as C18. The presence of the trityl group allowed for the desired full length oligonucleotide to be retained on the resin, while undesired truncated products that were acylated during the capping reaction were removed from the resin. These undesired oligonucleotides were removed from the resin with a lower percentage of an organic solvent, such as acetonitrile. After this removal of failure products, the trityl group was removed by acidic treatment (with an acid such as aqueous trifluoroacetic acid) while still on the C18 resin. After salt exchange and extensive washing of the resin with water, the desired deprotected product was eluted with a higher percentage of organic solvent in water. These purified oligonucleotides were then either lyophilized or annealed as required.

If the desired oligonucleotide contained any ribose (2′-hydroxyl) nucleotides, a modified procedure was required to remove the silyl 2′-hydroxyl protecting groups. After cleaving the oligonucleotide from the solid support with aqueous base, the column was further washed with an appropriate solvent, such as DMSO, to remove any material remaining on the column. After basic deprotection of the oligonucleotide, the reaction mixture then was treated with an appropriate fluoride reagent, such as triethylamine-hydrogen fluoride or TBAF, to cleave all of the silyl ethers and expose the desired alcohols. After silyl deprotection was complete, an appropriate buffer was added to each sample in order to neutralize the solution prior to purification (either by SAX or C18). In certain cases, a 5′-vinyl phosphonate-3′-phosphoramidite was coupled to the 5′-terminus of an oligonucleotide. The oxidation reagent for this incorporation may be changed to t-butyl hydroperoxide rather than iodine. When an oligonucleotide contained a protected phosphonate moiety, the methyl or ethyl phosphate esters were deprotected with an appropriate reagent, such as iodotrimethylsilane or bromotrimethylsilane in pyridine/acetonitrile, while the oligonucleotide was still on the solid support. After washing the support with acetonitrile, the oligonucleotide was treated with (3-mercaptoethanol in triethylamine/acetonitrile. After further washing with acetonitrile, the oligonucleotide product was cleaved from the solid support and fully deprotected with aqueous methylamine. The 5′-phosphonate oligonucleotides were purified by either SAX or C18 chromatography.

Each purified oligonucleotide was analyzed for purity by appropriate methods, including reversed phase HPLC, SAX HPLC, and capillary gel electrophoresis. The identity of the oligonucleotide was confirmed by mass spectrometry, using an ionization technique such as ESI or MALDI. The yields of each oligonucleotide were assessed by UV (260 nm) with a theoretically derived extinction coefficient. The appropriate amounts of each strand were approximated by UV (260 nm) measurements and theoretical EC260 extinction coefficients. The final oligonucleotide material was lyophilized or dissolved in an appropriate aqueous buffer prior to biochemical or biological evaluation.

Example 1

5′-HO;rG;rC;rU;rC;rA;rU;rU;rU;rA;rC;rC;rA;rG;rC;rG;rU;rA;rA;rA;rU;rG;rA;rG;rC-3′(SEQ ID NO. 838)

Oligosynthesis was conducted on a MerMade12 oligosynthesizer at 10 μmol scale. Suitably protected ribonucleoside 3′-phosphoramidites (5′-ODMT, 2′-OTBS, N-acetyl-cytidine, N-benzoyl-adenosine, N-isobutyryl-guanosine) were dissolved in acetonitrile (0.1 M). Ethylthio-tetrazole (ETT) was used as activator (0.25 M in acetonitrile). A solution of iodine (0.025 M) in pyridine/acetonitrile/water (45/45/10) was used as oxidizer. Detritylation was accomplished with a solution of 3% trichloroacetic acid (TCA) in methylene chloride. The CPG solid support was preloaded with the 3′-terminal nucleoside, 5′-ODMT-2′-OTBS-N-acetyl-cytidine. The DMT protecting group was removed with three cycles of 3% TCA, followed by 3×2 mL washes of the support with acetontrile. Equal volumes (0.5 mL each) of the required amidite solution (corresponding to the desired oligonucleotide sequence from the 3′-terminus) and activator was next added to the CPG. Nucleotide coupling was accomplished with three cycles of amidite/activator for 7.5 minutes each. After completion of the amidite coupling and washing of the CPG with acetonitrile (3×2 mL), the P(III) intermediate was oxidized to the P(V) phosphate ester with iodine (4×1 mL, 1 min each) and washed with acetonitrile (3×2 mL). Unreacted oligonucleotide 5′-hydroxyls were capped with an equal volume mixture (0.5 mL each) of 20% acetic anhydride/30% lutidine in acetonitrile and N-methyl-imidazole (20% in acetonitrile). The solid support was then washed with acetonitrile (4×2 mL), before the next coupling cycle was initiated with detritylation of the newly installed nucleotide unit.

After completion of the synthesis of the oligonucleotide sequence, the CPG support was washed with acetonitrile (4×2 mL). The protected oligonucleotide was removed from the solid support with 40% aqueous methylamine (3 mL, then 2 mL) and DMSO (2×3 mL). The combined solutions were heated at 35° C. for 45 minutes. The oligonucleotide was then treated with 3 mL of HF-triethylamine and heated to 40° C. for 60 minutes prior to the addition of 50 mM aqueous NaOAc (15 mL). The reaction mixture was directly loaded onto a C18 cartridge (5 g) that was prewashed with acetonitrile, water and 50 mM aqueous triethylammonium acetate (TEAA). After washing with water and 16% acetonitrile in 50 mM TEAA/50 mM NaCl (to remove shortmer synthesis failures), the DMT group was deprotected on-column with 1% TFA. The column was washed with water, 1 M NaCl and additional water prior to elution of the desired oligonucleotide with 30% aqueous acetonitrile.

The desired oligonucleotide was further purified by preparative mass-directed HPLC with a Waters X-Bridge Phenyl column (19×150 mm, ambient temperature, 25 mL/min, buffer A: 200 mM aqueous TEAA, pH 7, buffer B: 200 mM TEAA, 20% aq. acetonitrile, pH 7) eluting with a gradient of 3% to 11% B over 9 minutes and 11% to 25% B over 3 minutes. The counterion of the oligonucleotide was exchanged to sodium via spin filtration (MWCO 3000) and the isolated product was lyophilized to a white solid.

LC-MS: (m/z)⁻ (z=4) 1908.1 (exp); 1908.1 (theor); (z=5) 1526.1 (exp); 1526.3 (theor)

Examples 2-336, 342-352, 360-404, 436-442, 475-477, 514-593, 595-605, 607, 609-613, 615, 617-633, 635-645, 647, 649-653, 655, and 657-838 noted as SEQ ID No. 1 as shown in Table 5 below were prepared according to the procedures analogous to those outlined in Example 1. Examples 337-341, 353-359, 405-435, 443-474, 478-513, 594, 606, 608, 614, 616, 634, 646, 648, 654, and 656 with modified nucleobases in Table 5 below were prepared using procedures known in the art in combination with the procedures analogous to those outlined in Example 1.

TABLE 5 SEQ Ex Sequence MW ID No. 1 5′-HO;rG;rC;rU;rC;rA;rU;rU;rU;rA;rC;rC;rA;rG;rC;rG;rU; 7636.72 1 rA;rA;rA;rU;rG;rA;rG;rCSup-3′ 2 5′-HO;rG;fluC;rU;fluC;fluA;fluU;rU;fluU;rA;rC;fluC;rA; 7672.62 124 fluG;rC;fluG;fluU;fluA;rA;rA;rU;omeG;rA;rG;fluCSup-3′ 3 5′-HO;rG;fluC;rU;fluC;rA;fluU;omeU;rU;fluA;rC;rC;rA; 7694.68 125 fluG;rC;rG;rU;rA;omeA;fluA;fluU;rG;rA;fluG;omeCSup-3′ 4 5′-HO;rG;rC;fluU;rC;fluA;rU;rU;omeU;fluA;fluC;rC;rA; 7706.71 126 fluG;fluC;fluG;rU;rA;omeA;omeA;rU;rG;rA;rG;omeCSup-3′ 5 5′-HO;rG;rC;omeU;rC;rA;fluU;rU;omeU;fluA;rC;fluC;omeA; 7710.63 127 rG;rC;rG;fluU;rA;fluA;omeA;rU;fluG;fluA;fluG;fluCSup-3′ 6 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7716.81 128 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 7 5′-HO;rG;omeC;rU;fluC;rA;rU;fluU;omeU;fluA;omeC;fluC;fluA; 7720.73 129 rG;fluC;omeG;fluU;rA;fluA;fluA;fluU;fluG;fluA;fluG;fluCSup-3′ 8 5′-HO;rG;rC;fluU;rC;fluA;omeU;fluU;rU;rA;fluC;omeC;omeA; 7720.73 130 rG;fluC;omeG;omeU;rA;rA;rA;rU;fluG;rA;fluG;rCSup-3′ 9 5′-HO;rG;rC;rU;rC;fluA;omeU;omeU;fluU;rA;fluC;rC;fluA; 7722.70 131 rG;fluC;omeG;rU;rA;rA;fluA;fluU;omeG;fluA;rG;omeCSup-3′ 10 5′-HO;rG;rC;fluU;rC;fluA;fluU;fluU;omeU;fluA;omeC;rC;rA; 7724.64 132 omeG;omeC;fluG;rU;rA;fluA;omeA;fluU;fluG;rA;rG;rCSup-3′ 11 5′-HO;rG;rC;omeU;omeC;fluA;rU;rU;rU;fluA;fluC;fluC;rA; 7724.71 133 omeG;fluC;fluG;rU;omeA;rA;fluA;rU;fluG;fluA;omeG;rCSup-3′ 12 5′-HO;rG;rC;omeU;fluC;rA;rU;rU;rU;fluA;fluC;rC;fluA;fluG; 7724.68 134 rC;fluG;rU;rA;fluA;fluA;omeU;omeG;omeA;omeG;fluCSup-3′ 13 5′-HO;rG;omeC;fluU;rC;omeA;fluU;omeU;omeU;fluA;omeC; 7726.92 135 fluC;rA;fluG;fluC;rG;rU;fluA;rA;fluA;fluU;rG;fluA;rG;rCSup-3′ 14 5′-HO;rG;rC;fluU;fluC;fluA;omeU;fluU;rU;fluA;omeC;fluC;fluA; 7728.71 136 rG;fluC;rG;omeU;rA;rA;fluA;fluU;rG;omeA;omeG;fluCSup-3′ 15 5′-HO;rG;fluC;rU;omeC;rA;omeU;rU;omeU;omeA;fluC;rC;fluA; 7728.67 137 fluG;rC;fluG;omeU;fluA;rA;fluA;rU;fluG;fluA;fluG;fluCSup-3′ 16 5′-HO;rG;fluC;rU;rC;fluA;rU;fluU;fluU;fluA;rC;rC;omeA;fluG; 7730.58 138 fluC;omeG;fluU;omeA;omeA;fluA;omeU;fluG;fluA;fluG;rCSup-3′ 17 5′-HO;rG;omeC;fluU;omeC;rA;rU;rU;rU;rA;fluC;rC;rA;rG; 7730.78 139 rC;omeG;fluU;rA;fluA;fluA;omeU;omeG;omeA;rG;rCSup-3′ 18 5′-HO;rG;rC;omeU;rC;omeA;rU;fluU;fluU;rA;rC;omeC;omeA; 7730.75 140 rG;fluC;omeG;rU;rA;rA;rA;fluU;fluG;rA;omeG;rCSup-3′ 19 5′-HO;rG;rC;rU;rC;omeA;fluU;omeU;rU;rA;rC;fluC;omeA; 7732.73 141 omeG;rC;rG;fluU;rA;fluA;omeA;rU;rG;fluA;omeG;fluCSup-3′ 20 5′-HO;rG;rC;rU;fluC;fluA;fluU;omeU;rU;rA;fluC;omeC;rA;rG; 7734.77 142 omeC;rG;omeU;fluA;omeA;rA;omeU;fluG;rA;fluG;rCSup-3′ 21 5′-HO;rG;omeC;fluU;rC;fluA;fluU;fluU;rU;rA;fluC;rC;rA;rG; 7736.75 143 fluC;rG;omeU;omeA;omeA;fluA;rU;omeG;rA;omeG;fluCSup-3′ 22 5′-HO;rG;rC;rU;rC;fluA;rU;fluU;rU;fluA;omeC;fluC;omeA;rG; 7736.71 144 fluC;omeG;fluU;fluA;rA;omeA;omeU;omeG;rA;fluG;rCSup-3′ 23 5′-HO;rG;omeC;fluU;rC;omeA;omeU;rU;fluU;omeA;rC;fluC;rA; 7736.68 145 fluG;rC;fluG;omeU;fluA;fluA;rA;omeU;fluG;rA;rG;rCSup-3′ 24 5′-HO;rG;omeC;rU;rC;rA;rU;fluU;rU;omeA;rC;omeC;omeA;fluG; 7736.71 146 fluC;rG;fluU;rA;omeA;fluA;fluU;fluG;fluA;omeG;rCSup-3′ 25 5′-HO;rG;omeC;omeU;rC;fluA;fluU;fluU;omeU;rA;omeC;omeC; 7736.75 147 rA;rG;rC;fluG;fluU;rA;fluA;fluA;omeU;rG;rA;rG;fluCSup-3′ 26 5′-HO;rG;rC;fluU;omeC;omeA;fluU;rU;fluU;rA;omeC;fluC;rA; 7736.75 148 omeG;rC;rG;rU;rA;rA;fluA;fluU;omeG;omeA;fluG;fluCSup-3′ 27 5′-HO;rG;omeC;fluU;rC;rA;omeU;fluU;fluU;fluA;omeC;rC;rA; 7738.73 149 rG;fluC;omeG;omeU;fluA;fluA;fluA;rU;fluG;rA;rG;omeCSup-3′ 28 5′-HO;rG;omeC;omeU;fluC;fluA;rU;fluU;rU;rA;rC;rC;fluA;omeG; 7738.69 150 rC;rG;fluU;omeA;fluA;fluA;fluU;omeG;fluA;rG;omeCSup-3′ 29 5′-HO;rG;fluC;fluU;rC;fluA;omeU;rU;rU;fluA;rC;rC;fluA;rG; 7738.69 151 omeC;omeG;omeU;fluA;fluA;omeA;rU;fluG;fluA;rG;omeCSup-3′ 30 5′-HO;rG;fluC;omeU;rC;omeA;fluU;omeU;fluU;fluA;omeC;omeC; 7740.67 152 rA;fluG;rC;fluG;rU;omeA;fluA;fluA;rU;fluG;rA;fluG;rCSup-3′ 31 5′-HO;rG;fluC;fluU;omeC;rA;fluU;fluU;rU;rA;fluC;fluC;omeA; 7742.75 153 omeG;omeC;fluG;fluU;rA;rA;fluA;omeU;fluG;fluA;rG;omeCSup-3′ 32 5′-HO;rG;rC;rU;omeC;omeA;fluU;fluU;fluU;omeA;omeC;fluC; 7746.77 154 fluA;rG;rC;rG;omeU;rA;omeA;rA;omeU;rG;fluA;rG;rCSup-3′ 33 5′-HO;rG;rC;omeU;omeC;rA;omeU;fluU;rU;omeA;rC;fluC;rA; 7746.81 155 omeG;fluC;omeG;rU;rA;fluA;rA;rU;fluG;rA;omeG;fluCSup-3′ 34 5′-HO;rG;omeC;rU;omeC;omeA;omeU;omeU;fluU;fluA;fluC;rC; 7748.82 156 fluA;rG;fluC;rG;rU;rA;rA;omeA;rU;rG;fluA;omeG;fluCSup-3′ 35 5′-HO;rG;fluC;rU;omeC;fluA;omeU;omeU;fluU;rA;fluC;fluC;rA; 7750.76 157 rG;rC;omeG;fluU;omeA;rA;fluA;fluU;rG;omeA;omeG;rCSup-3′ 36 5′-HO;rG;fluC;fluU;omeC;rA;rU;fluU;omeU;fluA;rC;rC;rA;fluG; 7750.76 158 fluC;omeG;rU;fluA;omeA;rA;rU;omeG;omeA;omeG;fluCSup-3′ 37 5′-HO;rG;rC;fluU;omeC;omeA;rU;fluU;fluU;rA;omeC;fluC;fluA; 7752.74 159 fluG;rC;omeG;fluU;omeA;rA;fluA;omeU;rG;rA;fluG;omeCSup-3′ 38 5′-HO;rG;omeC;omeU;fluC;omeA;rU;fluU;rU;fluA;omeC;rC;rA;rG; 7752.77 160 omeC;fluG;fluU;fluA;rA;fluA;rU;fluG;fluA;omeG;omeCSup-3′ 39 5′-HO;rG;rC;rU;rC;rA;rU;omeU;omeU;rA;fluC;fluC;fluA;omeG; 7752.74 161 fluC;omeG;fluU;fluA;omeA;fluA;fluU;omeG;rA;omeG;fluCSup-3′ 40 5′-HO;rG;rC;fluU;rC;rA;rU;fluU;fluU;omeA;omeC;rC;omeA;fluG; 7752.67 162 rC;fluG;omeU;fluA;omeA;fluA;fluU;rG;omeA;fluG;omeCSup-3′ 41 5′-HO;rG;fluC;omeU;omeC;omeA;rU;fluU;omeU;rA;fluC;rC;rA; 7754.75 163 omeG;fluC;rG;omeU;rA;fluA;omeA;fluU;fluG;fluA;fluG;fluCSup-3′ 42 5′-HO;rG;rC;omeU;omeC;fluA;fluU;fluU;rU;rA;fluC;fluC;omeA; 7756.73 164 fluG;fluC;omeG;rU;fluA;fluA;rA;fluU;omeG;fluA;omeG;omeCSup-3′ 43 5′-HO;rG;omeC;rU;rC;omeA;fluU;fluU;fluU;rA;omeC;fluC;omeA; 7756.73 165 rG;omeC;fluG;rU;omeA;fluA;fluA;fluU;fluG;omeA;fluG;fluCSup-3′ 44 5′-HO;rG;rC;fluU;omeC;rA;omeU;rU;omeU;omeA;rC;omeC; 7756.83 166 rA;rG;rC;fluG;rU;omeA;omeA;omeA;rU;rG;rA;fluG;fluCSup-3′ 45 5′-HO;rG;rC;rU;fluC;rA;rU;rU;omeU;omeA;omeC;omeC;rA; 7756.87 167 fluG;fluC;omeG;omeU;rA;rA;fluA;omeU;rG;rA;omeG;rCSup-3′ 46 5′-HO;rG;omeC;fluU;fluC;omeA;fluU;rU;fluU;omeA;fluC;fluC; 7758.75 168 omeA;fluG;fluC;rG;omeU;omeA;rA;fluA;omeU;fluG;rA;fluG;fluCSup-3′ 47 5′-HO;rG;rC;omeU;fluC;rA;rU;rU;rU;rA;rC;omeC;fluA;fluG;fluC; 7760.82 169 rG;omeU;fluA;omeA;omeA;omeU;fluG;omeA;rG;omeCSup-3′ 48 5′-HO;rG;fluC;rU;rC;rA;omeU;omeU;fluU;fluA;rC;fluC;omeA; 7760.79 170 omeG;rC;fluG;omeU;rA;rA;omeA;rU;omeG;rA;fluG;omeCSup-3′ 49 5′-HO;rG;rC;rU;fluC;omeA;omeU;fluU;rU;fluA;rC;rC;fluA;omeG; 7762.76 171 omeC;rG;rU;fluA;rA;omeA;omeU;fluG;omeA;omeG;fluCSup-3′ 50 5′-HO;rG;fluC;rU;omeC;omeA;fluU;omeU;rU;omeA;rC;rC;rA;fluG; 7762.76 172 fluC;rG;omeU;fluA;fluA;omeA;omeU;omeG;fluA;rG;rCSup-3′ 51 5′-HO;rG;omeC;rU;omeC;omeA;omeU;rU;omeU;fluA;omeC;fluC; 7762.87 173 rA;fluG;fluC;rG;fluU;fluA;rA;omeA;fluU;rG;rA;rG;omeCSup-3′ 52 5′-HO;rG;fluC;fluU;omeC;omeA;rU;omeU;rU;rA;rC;fluC;fluA;rG; 7764.81 174 omeC;fluG;omeU;rA;omeA;fluA;rU;omeG;fluA;omeG;fluCSup-3′ 53 5′-HO;rG;fluC;omeU;fluC;fluA;rU;rU;omeU;rA;fluC;fluC;omeA;rG; 7764.85 175 fluC;omeG;rU;omeA;rA;omeA;omeU;rG;fluA;omeG;fluCSup-3′ 54 5′-HO;rG;fluC;fluU;fluC;omeA;omeU;fluU;rU;rA;fluC;omeC;rA; 7766.79 176 fluG;rC;omeG;omeU;rA;fluA;omeA;fluU;rG;fluA;omeG;omeCSup-3′ 55 5′-HO;rG;fluC;fluU;fluC;rA;fluU;fluU;rU;omeA;fluC;fluC;omeA; 7768.80 177 omeG;omeC;rG;fluU;omeA;fluA;rA;rU;omeG;omeA;fluG;omeCSup-3′ 56 5′-HO;rG;omeC;rU;fluC;omeA;fluU;fluU;omeU;fluA;rC;omeC;fluA; 7768.77 178 rG;fluC;fluG;omeU;fluA;rA;omeA;omeU;rG;omeA;fluG;fluCSup-3′ 57 5′-HO;rG;omeC;fluU;rC;fluA;omeU;rU;fluU;fluA;rC;omeC;fluA; 7768.73 179 omeG;fluC;fluG;rU;omeA;fluA;fluA;fluU;omeG;rA;omeG;omeCSup-3′ 58 5′-HO;rG;rC;fluU;fluC;omeA;fluU;omeU;omeU;omeA;fluC;rC;omeA; 7770.71 180 fluG;omeC;omeG;fluU;fluA;fluA;fluA;fluU;rG;omeA;rG;fluCSup-3′ 59 5′-HO;rG;omeC;fluU;rC;fluA;fluU;omeU;omeU;omeA;omeC;fluC; 7770.71 181 fluA;fluG;rC;omeG;fluU;fluA;fluA;rA;rU;omeG;fluA;omeG;fluCSup-3′ 60 5′-HO;rG;fluC;omeU;omeC;fluA;omeU;fluU;fluU;fluA;fluC;omeC;fluA; 7772.76 182 fluG;omeC;omeG;fluU;rA;omeA;rA;omeU;fluG;fluA;rG;fluCSup-3′ 61 5′-HO;rG;fluC;omeU;rC;fluA;omeU;rU;omeU;rA;rC;omeC;fluA;rG; 7776.81 183 omeC;rG;fluU;omeA;omeA;rA;fluU;fluG;omeA;fluG;omeCSup-3′ 62 5′-HO;rG;fluC;rU;rC;fluA;fluU;omeU;omeU;fluA;fluC;omeC;omeA; 7776.81 184 omeG;rC;omeG;omeU;rA;rA;fluA;rU;fluG;rA;omeG;omeCSup-3′ 63 5′-HO;rG;omeC;omeU;omeC;rA;rU;fluU;fluU;rA;rC;omeC;omeA; 7776.85 185 rG;omeC;rG;omeU;fluA;fluA;fluA;fluU;omeG;rA;omeG;fluCSup-3′ 64 5′-HO;rG;omeC;fluU;omeC;rA;omeU;rU;omeU;fluA;omeC;fluC;fluA; 7776.81 186 fluG;rC;rG;omeU;rA;rA;omeA;fluU;omeG;fluA;omeG;rCSup-3′ 65 5′-HO;rG;rC;rU;fluC;omeA;fluU;rU;rU;fluA;omeC;omeC;rA;omeG; 7776.85 187 omeC;omeG;omeU;rA;fluA;fluA;omeU;fluG;omeA;rG;fluCSup-3′ 66 5′-HO;rG;fluC;omeU;fluC;rA;omeU;rU;fluU;rA;omeC;omeC;omeA; 7778.82 188 rG;rC;omeG;fluU;fluA;fluA;fluA;omeU;fluG;omeA;rG;omeCSup-3′ 67 5′-HO;rG;fluC;omeU;fluC;fluA;fluU;fluU;rU;rA;rC;omeC;omeA;fluG; 7778.82 189 fluC;omeG;rU;rA;omeA;omeA;omeU;omeG;fluA;rG;omeCSup-3′ 68 5′-HO;rG;omeC;fluU;fluC;fluA;omeU;omeU;rU;omeA;rC;omeC;omeA; 7778.79 190 omeG;fluC;omeG;rU;fluA;fluA;fluA;omeU;rG;fluA;rG;rCSup-3′ 69 5′-HO;rG;rC;omeU;fluC;omeA;omeU;rU;rU;fluA;fluC;fluC;omeA; 7778.79 191 omeG;rC;rG;fluU;fluA;omeA;fluA;omeU;rG;omeA;omeG;fluCSup-3′ 70 5′-HO;rG;omeC;rU;fluC;rA;omeU;fluU;rU;fluA;fluC;omeC;rA;omeG; 7780.80 192 rC;omeG;omeU;fluA;omeA;omeA;fluU;fluG;omeA;fluG;fluCSup-3′ 71 5′-HO;rG;rC;omeU;fluC;rA;omeU;omeU;fluU;fluA;fluC;omeC;fluA; 7780.80 193 fluG;omeC;fluG;fluU;rA;fluA;omeA;rU;rG;omeA;omeG;omeCSup-3′ 72 5′-HO;rG;omeC;omeU;fluC;rA;fluU;rU;rU;omeA;fluC;fluC;fluA;rG; 7780.80 194 omeC;fluG;omeU;omeA;omeA;fluA;omeU;fluG;fluA;omeG;rCSup-3′ 73 5′-HO;rG;fluC;omeU;rC;omeA;omeU;rU;fluU;rA;omeC;omeC;omeA; 7782.78 195 fluG;fluC;fluG;rU;fluA;fluA;omeA;fluU;omeG;omeA;fluG;fluCSup-3′ 74 5′-HO;rG;fluC;fluU;omeC;fluA;fluU;omeU;rU;fluA;omeC;rC;fluA;fluG; 7784.76 196 omeC;rG;fluU;omeA;omeA;fluA;omeU;fluG;omeA;omeG;fluCSup-3′ 75 5′-HO;rG;omeC;fluU;fluC;fluA;omeU;fluU;fluU;omeA;fluC;omeC;fluA; 7784.75 197 omeG;rC;fluG;fluU;omeA;rA;omeA;rU;omeG;fluA;fluG;omeCSup-3′ 76 5′-HO;rG;omeC;rU;omeC;rA;omeU;fluU;fluU;rA;rC;fluC;omeA; 7784.89 198 fluG;omeC;omeG;rU;omeA;rA;omeA;rU;omeG;omeA;rG;rCSup-3′ 77 5′-HO;rG;rC;rU;omeC;omeA;fluU;rU;rU;omeA;omeC;omeC;fluA; 7786.90 199 rG;omeC;omeG;rU;fluA;rA;fluA;omeU;rG;omeA;fluG;omeCSup-3′ 78 5′-HO;rG;fluC;omeU;fluC;fluA;omeU;omeU;omeU;omeA;omeC; 7786.91 200 omeC;fluA;omeG;fluC;rG;rU;rA;omeA;rA;rU;omeG;rA;rG;rCSup-3′ 79 5′-HO;rG;rC;rU;fluC;omeA;omeU;fluU;omeU;fluA;omeC;rC;rA;fluG; 7786.83 201 omeC;omeG;rU;omeA;omeA;fluA;rU;omeG;rA;omeG;rCSup-3′ 80 5′-HO;rG;fluC;omeU;fluC;omeA;fluU;omeU;omeU;rA;rC;rC;fluA;fluG; 7792.77 202 rC;omeG;omeU;omeA;fluA;fluA;rU;fluG;omeA;omeG;omeCSup-3′ 81 5′-HO;rG;omeC;omeU;omeC;fluA;omeU;rU;rU;omeA;fluC;omeC;fluA; 7794.85 203 fluG;omeC;fluG;rU;fluA;omeA;omeA;fluU;fluG;rA;omeG;fluCSup-3′ 82 5′-HO;rG;fluC;omeU;rC;omeA;omeU;fluU;rU;fluA;omeC;omeC;fluA; 7794.82 204 fluG;fluC;fluG;omeU;omeA;omeA;rA;fluU;omeG;omeA;rG;fluCSup-3′ 83 5′-HO;rG;rC;fluU;omeC;fluA;omeU;rU;fluU;omeA;omeC;fluC;rA;omeG; 7794.81 205 fluC;fluG;omeU;fluA;fluA;rA;omeU;fluG;omeA;omeG;omeCSup-3′ 84 5′-HO;rG;rC;omeU;rC;omeA;omeU;fluU;fluU;rA;rC;rC;omeA;omeG; 7798.80 206 rC;omeG;omeU;omeA;rA;fluA;omeU;omeG;fluA;rG;omeCSup-3′ 85 5′-HO;rG;omeC;omeU;rC;rA;fluU;rU;rU;fluA;rC;fluC;omeA;omeG; 7798.90 207 omeC;omeG;omeU;omeA;rA;rA;fluU;rG;omeA;omeG;omeCSup-3′ 86 5′-HO;rG;omeC;omeU;omeC;omeA;fluU;omeU;omeU;fluA;fluC;fluC; 7802.90 208 omeA;rG;omeC;omeG;fluU;fluA;omeA;omeA;rU;rG;rA;rG;rCSup-3′ 87 5′-HO;rG;omeC;fluU;fluC;omeA;rU;omeU;fluU;fluA;omeC;fluC;omeA; 7802.93 209 fluG;omeC;rG;omeU;omeA;rA;omeA;rU;omeG;rA;rG;omeCSup-3′ 88 5′-HO;rG;rC;omeU;omeC;rA;fluU;omeU;fluU;rA;omeC;omeC;rA;omeG; 7802.90 210 fluC;omeG;omeU;omeA;omeA;omeA;fluU;rG;fluA;rG;fluCSup-3′ 89 5′-HO;rG;rC;fluU;omeC;omeA;rU;fluU;omeU;fluA;omeC;omeC;omeA; 7816.87 211 rG;rC;omeG;fluU;omeA;omeA;rA;omeU;fluG;fluA;omeG;omeCSup-3′ 90 5′-HO;rG;omeC;fluU;omeC;omeA;omeU;rU;rU;fluA;fluC;omeComeA; 7820.86 212 omeG;fluC;omeG;omeU;omeA;fluA;fluA;fluU;omeG;omeA;fluG;rCSup-3′ Exact 7817.20 91 5′-HO;rG;rC;omeU;omeC;rA;fluU;omeU;omeU;fluA;omeC;rC;omeA; 7856.93 213 omeG;omeC;omeG;omeU;omeA;omeA;omeA;omeU;omeG;fluA;fluG; fluCSup-3′ 92 5′-HO;rG;fluC;fluU;fluC;fluA;fluU;fluU;fluU;fluA;fluC;fluC;fluA;fluG; 7682.40 214 fluC;fluG;fluU;fluA;fluA;fluA;fluU;fluG;fluA;fluG;fluCSup-3′ 93 5′-HO;rG;rC;fluU;rC;rA;fluU;rU;omeU;rA;rC;omeC;fluA; 7692.66 215 omeG;fluC;rG;fluU;fluA;rA;rA;rU;rG;fluA;rG;rCSup-3′ 94 5′-HO;rG;rC;fluU;rC;rA;fluU;fluU;rU;omeA;rC;fluC;fluA; 7696.58 216 rG;rC;fluG;rU;fluA;omeA;rA;fluU;omeG;fluA;rG;rCSup-3′ 95 5′-HO;rG;rC;fluU;rC;rA;rU;omeU;rU;fluA;fluC;omeC;rA;rG; 7706.71 217 rC;rG;fluU;omeA;rA;rA;fluU;omeG;fluA;rG;fluCSup-3′ 96 5′-HO;rG;fluC;omeU;fluC;rA;fluU;omeU;rU;rA;fluC;fluC;fluA; 7710.70 218 rG;rC;rG;rU;omeA;rA;fluA;fluU;omeG;rA;fluG;rCSup-3′ 97 5′-HO;rG;fluC;rU;rC;fluA;rU;rU;rU;rA;fluC;fluC;omeA;omeG; 7712.72 219 fluC;fluG;omeU;fluA;omeA;rA;fluU;rG;fluA;rG;fluCSup-3′ 98 5′-HO;rG;rC;fluU;fluC;omeA;fluU;fluU;fluU;fluA;rC;fluC;fluA; 7718.58 220 rG;fluC;fluG;fluU;rA;omeA;omeA;fluU;fluG;omeA;rG;rCSup-3′ 99 5′-HO;rG;rC;omeU;rC;rA;omeU;fluU;fluU;fluA;fluC;rC;rA;fluG; 7720.69 221 rC;omeG;rU;omeA;rA;rA;omeU;rG;fluA;rG;fluCSup-3′ 100 5′-HO;rG;fluC;fluU;fluC;rA;fluU;fluU;rU;rA;omeC;omeC;rA; 7724.79 222 omeG;fluC;fluG;rU;rA;rA;rA;rU;omeG;omeA;fluG;fluCSup-3′ 101 5′-HO;rG;omeC;rU;fluC;fluA;fluU;fluU;fluU;fluA;fluC;rC;omeA; 7724.68 223 fluG;rC;rG;omeU;rA;fluA;rA;omeU;rG;omeA;rG;rCSup-3′ 102 5′-HO;rG;fluC;rU;omeC;fluA;fluU;rU;rU;omeA;rC;rC;omeA;rG; 7728.67 224 fluC;fluG;fluU;rA;fluA;fluA;fluU;fluG;fluA;omeG;omeCSup-3′ 103 5′-HO;rG;rC;rU;omeC;fluA;fluU;rU;fluU;rA;fluC;rC;rA;rG; 7732.79 225 omeC;rG;fluU;fluA;rA;rA;rU;omeG;omeA;omeG;omeCSup-3′ 104 5′-HO;rG;omeC;omeU;rC;rA;rU;omeU;rU;fluA;rC;rC;fluA; 7732.73 226 omeG;fluC;rG;omeU;fluA;fluA;rA;omeU;rG;rA;fluG;rCSup-3′ 105 5′-HO;rG;omeC;fluU;rC;rA;rU;fluU;rU;rA;fluC;rC;omeA;rG; 7734.77 227 omeC;fluG;rU;fluA;omeA;rA;omeU;rG;omeA;fluG;fluCSup-3′ 106 5′-HO;rG;fluC;rU;rC;fluA;rU;rU;omeU;fluA;rC;fluC;rA;fluG; 7734.77 228 omeC;rG;rU;rA;fluA;rA;omeU;omeG;omeA;fluG;omeCSup-3′ 107 5′-HO;rG;fluC;fluU;fluC;fluA;rU;rU;omeU;omeA;rC;rC;omeA; 7734.77 229 rG;omeC;fluG;rU;omeA;rA;rA;rU;rG;fluA;omeG;fluCSup-3′ 108 5′-HO;rG;fluC;fluU;omeC;rA;rU;fluU;rU;omeA;omeC;fluC;rA; 7736.82 230 fluG;fluC;omeG;fluU;omeA;omeA;rA;rU;rG;rA;rG;fluCSup-3′ 109 5′-HO;rG;rC;fluU;omeC;omeA;rU;fluU;omeU;rA;fluC;omeC; 7736.75 231 fluA;omeG;omeC;rG;rU;rA;fluA;fluA;rU;rG;fluA;fluG;rCSup-3′ 110 5′-HO;rG;fluC;rU;rC;omeA;fluU;fluU;fluU;fluA;rC;omeC;rA;rG; 7736.75 232 omeC;rG;rU;omeA;fluA;rA;fluU;rG;fluA;omeG;omeCSup-3′ 111 5′-HO;rG;rC;rU;fluC;rA;rU;rU;omeU;fluA;fluC;fluC;fluA;rG; 7736.79 233 omeC;fluG;rU;omeA;fluA;fluA;omeU;omeG;rA;rG;omeCSup-3′ 112 5′-HO;rG;fluC;rU;fluC;omeA;rU;rU;omeU;fluA;fluC;omeC; 7736.75 234 omeA;rG;rC;fluG;fluU;fluA;omeA;fluA;rU;omeG;rA;rG;rCSup-3′ 113 5′-HO;rG;rC;rU;rC;omeA;rU;fluU;fluU;omeA;fluC;fluC;rA;fluG; 7738.73 235 omeC;rG;fluU;omeA;omeA;omeA;fluU;fluG;rA;rG;fluCSup-3′ 114 5′-HO;rG;rC;rU;fluC;fluA;rU;omeU;fluU;rA;omeC;rC;fluA;omeG; 7738.69 236 rC;fluG;rU;fluA;omeA;omeA;rU;fluG;fluA;fluG;omeCSup-3′ 115 5′-HO;rG;omeC;fluU;fluC;fluA;fluU;rU;fluU;omeA;rC;fluC;fluA; 7738.73 237 rG;rC;omeG;rU;rA;omeA;omeA;omeU;fluG;rA;rG;fluCSup-3′ 116 5′-HO;rG;omeC;rU;rC;rA;omeU;rU;fluU;rA;fluC;rC;fluA;fluG; 7738.69 238 fluC;omeG;omeU;omeA;omeA;fluA;rU;fluG;fluA;fluG;rCSup-3′ 117 5′-HO;rG;omeC;fluU;fluC;rA;rU;omeU;fluU;omeA;omeC;fluC;fluA; 7740.78 239 fluG;fluC;rG;rU;omeA;fluA;rA;rU;fluG;fluA;rG;omeCSup-3′ 118 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7744.80 240 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 119 5′-HO;rG;fluC;rU;fluC;rA;omeU;fluU;fluU;fluA;fluC;fluC;rA;omeG; 7746.71 241 omeC;fluG;omeU;fluA;omeA;fluA;fluU;rG;fluA;fluG;omeCSup-3′ 120 5′-HO;rG;rC;fluU;omeC;rA;omeU;fluU;omeU;omeA;fluC;rC; 7746.77 242 omeA;fluG;rC;rG;omeU;rA;rA;rA;fluU;fluG;rA;rG;omeCSup-3′ 121 5′-HO;rG;fluC;fluU;rC;omeA;rU;rU;rU;rA;omeC;omeC;fluA;fluG; 7750.76 243 omeC;omeG;omeU;fluA;rA;omeA;fluU;fluG;rA;fluG;rCSup-3′ 122 5′-HO;rG;fluC;omeU;omeC;fluA;fluU;omeU;fluU;fluA;omeC;rC;fluA; 7752.74 244 rG;fluC;rG;omeU;omeA;fluA;rA;fluU;rG;rA;omeG;rCSup-3′ 123 5′-HO;rG;rC;omeU;rC;fluA;omeU;fluU;omeU;omeA;rC;fluC;rA;fluG; 7752.67 245 omeC;fluG;fluU;fluA;rA;fluA;omeU;rG;fluA;omeG;rCSup-3′ 124 5′-HO;rG;fluC;omeU;fluC;rA;rU;rU;fluU;omeA;omeC;rC;omeA;rG; 7752.74 246 fluC;fluG;omeU;fluA;fluA;omeA;fluU;rG;fluA;omeG;rCSup-3′ 125 5′-HO;rG;fluC;omeU;omeC;omeA;rU;fluU;fluU;omeA;fluC;fluC;fluA; 7752.78 247 rG;omeC;rG;rU;fluA;fluA;omeA;fluU;rG;omeA;rG;rCSup-3′ 126 5′-HO;rG;omeC;rU;fluC;rA;rU;omeU;rU;omeA;fluC;omeC;rA;fluG; 7754.79 248 fluC;omeG;fluU;fluA;fluA;rA;omeU;omeG;fluA;fluG;fluCSup-3′ 127 5′-HO;rG;omeC;omeU;omeC;fluA;fluU;rU;omeU;fluA;fluC;rC;omeA; 7754.68 249 fluG;rC;fluG;fluU;rA;fluA;rA;fluU;omeG;omeA;fluG;rCSup-3′ 128 5′-HO;rG;fluC;rU;omeC;rA;omeU;rU;rU;omeA;rC;omeC;fluA; 7754.86 250 omeG;rC;rG;rU;omeA;omeA;rA;fluU;rG;omeA;rG;rCSup-3′ 129 5′-HO;rG;omeC;fluU;fluC;rA;fluU;omeU;omeU;rA;rC;fluC;fluA;fluG; 7756.73 251 omeC;omeG;fluU;fluA;rA;rA;fluU;fluG;fluA;omeG;omeCSup-3′ 130 5′-HO;rG;omeC;omeU;omeC;omeA;rU;rU;omeU;fluA;rC;rC; 7756.83 252 fluA;omeG;rC;omeG;rU;fluA;rA;rA;rU;fluG;rA;rG;omeCSup-3′ 131 5′-HO;rG;omeC;omeU;fluC;omeA;rU;rU;fluU;rA;rC;rC;omeA; 7756.83 253 omeG;rC;rG;fluU;rA;omeA;fluA;rU;rG;rA;omeG;omeCSup-3′ 132 5′-HO;rG;rC;rU;rC;omeA;rU;rU;fluU;omeA;fluC;fluC;omeA;rG; 7758.85 254 fluC;rG;omeU;rA;omeA;fluA;rU;omeG;omeA;rG;omeCSup-3′ 133 5′-HO;rG;rC;fluU;rC;rA;omeU;omeU;omeU;rA;omeC;rC;omeA; 7758.81 255 rG;omeC;rG;rU;fluA;omeA;fluA;omeU;rG;rA;fluG;fluCSup-3′ 134 5′-HO;rG;fluC;omeU;fluC;omeA;omeU;rU;omeU;rA;rC;omeC;rA; 7760.86 256 rG;fluC;fluG;rU;omeA;rA;rA;omeU;fluG;fluA;rG;omeCSup-3′ 135 5′-HO;rG;fluC;rU;rC;rA;omeU;omeU;omeU;rA;omeC;fluC;omeA;rG; 7760.79 257 rC;omeG;rU;fluA;fluA;fluA;rU;fluG;omeA;omeG;rCSup-3′ 136 5′-HO;rG;fluC;fluU;rC;omeA;rU;omeU;fluU;omeA;fluC;rC;rA;omeG; 7760.75 258 rC;rG;rU;fluA;rA;omeA;omeU;fluG;omeA;omeG;rCSup-3′ 137 5′-HO;rG;fluC;omeU;rC;rA;omeU;rU;fluU;fluA;fluC;fluC;rA;fluG;omeC; 7762.80 259 omeG;fluU;rA;rA;omeA;omeU;rG;omeA;omeG;rCSup-3′ 138 5′-HO;rG;fluC;omeU;rC;omeA;rU;omeU;rU;omeA;omeC;fluC;fluA; 7762.84 260 omeG;fluC;fluG;fluU;rA;rA;rA;rU;rG;omeA;fluG;omeCSup-3′ 139 5′-HO;rG;rC;fluU;omeC;fluA;rU;omeU;rU;fluA;rC;omeC;rA;fluG;rC; 7762.73 261 fluG;omeU;omeA;rA;omeA;fluU;omeG;fluA;omeG;rCSup-3′ 140 5′-HO;rG;fluC;fluU;rC;rA;rU;rU;fluU;omeA;rC;omeC;fluA;omeG;omeC; 7763.80 262 omeG;fluU;rA;rA;omeA;omeU;fluG;rA;omeG;fluCSup-3′ 141 5′-HO;rG;rC;fluU;omeC;rA;omeU;fluU;fluU;omeA;omeC;omeC;rA; 7764.77 263 rG;omeC;rG;fluU;fluA;fluA;omeA;rU;omeG;fluA;fluG;rCSup-3′ 142 5′-HO;rG;rC;rU;rC;omeA;omeU;omeU;rU;omeA;fluC;rC;omeA;omeG; 7764.74 264 fluC;fluG;fluU;omeA;fluA;fluA;rU;fluG;omeA;rG;fluCSup-3′ 143 5′-HO;rG;rC;omeU;fluC;rA;fluU;omeU;fluU;omeA;rC;rC;rA;omeG; 7766.68 265 fluC;fluG;fluU;fluA;omeA;fluA;fluU;omeG;omeA;omeG;rCSup-3′ 144 5′-HO;rG;fluC;fluU;omeC;omeA;omeU;fluU;rU;rA;rC;rC;rA;omeG; 7766.75 266 fluC;fluG;fluU;omeA;omeA;omeA;omeU;rG;fluA;fluG;fluCSup-3′ 145 5′-HO;rG;omeC;rU;omeC;fluA;omeU;omeU;fluU;rA;omeC;fluC;fluA; 7766.82 267 omeG;omeC;fluG;omeU;rA;fluA;rA;fluU;fluG;rA;rG;fluCSup-3′ 146 5′-HO;rG;omeC;rU;omeC;fluA;fluU;fluU;fluU;omeA;fluC;omeC;omeA; 7768.80 268 fluG;fluC;rG;rU;fluA;rA;omeA;rU;fluG;omeA;fluG;omeCSup-3′ 147 5′-HO;rG;omeC;omeU;fluC;omeA;fluU;rU;fluU;fluA;omeC;omeC;rA; 7770.78 269 omeG;omeC;omeG;fluU;fluA;fluA;rA;fluU;rG;fluA;fluG;fluCSup-3′ 148 5′-HO;rG;fluC;fluU;fluC;omeA;omeU;omeU;omeU;fluA;fluC;omeC; 7770.78 270 fluA;rG;fluC;rG;fluU;omeA;fluA;rA;omeU;omeG;rA;fluG;fluCSup-3′ 149 5′-HO;rG;omeC;fluU;rC;fluA;fluU;rU;rU;omeA;omeC;omeC;omeA; 7770.88 271 omeG;omeC;omeG;rU;omeA;rA;fluA;rU;rG;rA;rG;rCSup-3′ 150 5′-HO;rG;rC;rU;fluC;rA;omeU;fluU;omeU;rA;rC;rC;omeA;omeG;fluC; 7770.81 272 omeG;fluU;omeA;rA;rA;omeU;omeG;rA;omeG;rCSup-3′ 151 5′-HO;rG;omeC;omeU;omeC;rA;rU;omeU;omeU;rA;rC;fluC;omeA; 7772.82 273 omeG;rC;fluG;rU;fluA;fluA;omeA;rU;fluG;omeA;rG;rCSup-3′ 152 5′-HO;rG;omeC;fluU;fluC;omeA;omeU;rU;omeU;rA;omeC;rC;omeA; 7772.82 274 fluG;rC;fluG;fluU;omeA;omeA;rA;rU;rG;omeA;rG;rCSup-3′ 153 5′-HO;rG;fluC;rU;fluC;fluA;rU;rU;omeU;omeA;omeC;rC;omeA;rG; 7774.87 275 omeC;omeG;omeU;rA;rA;omeA;fluU;omeG;fluA;rG;fluCSup-3′ 154 5′-HO;rG;rC;rU;omeC;omeA;rU;omeU;rU;fluA;rC;fluC;omeA;rG;rC; 7774.80 276 omeG;fluU;omeA;fluA;omeA;fluU;omeG;rA;fluG;omeCSup-3′ 155 5′-HO;rG;fluC;rU;rC;rA;rU;fluU;omeU;rA;omeC;fluC;fluA;fluG;rC; 7776.81 277 rG;omeU;omeA;fluA;omeA;omeU;omeG;omeA;omeG;fluCSup-3′ 156 5′-HO;rG;fluC;fluU;omeC;rA;rU;rU;fluU;fluA;omeC;omeC;omeA;omeG; 7776.81 278 rC;rG;fluU;omeA;omeA;rA;omeU;fluG;fluA;omeG;rCSup-3′ 157 5′-HO;rG;fluC;omeU;fluC;rA;omeU;fluU;omeU;omeA;fluC;fluC;omeA; 7778.83 279 omeG;omeC;rG;rU;rA;fluA;omeA;fluU;fluG;rA;omeG;rCSup-3′ 158 5′-HO;rG;rC;fluU;omeC;fluA;omeU;omeU;omeU;fluA;rC;omeC;omeA; 7778.75 280 fluG;omeC;fluG;fluU;rA;omeA;rA;fluU;rG;omeA;fluG;rCSup-3′ 159 5′-HO;rG;rC;omeU;fluC;fluA;omeU;fluU;fluU;fluA;omeC;fluC;omeA; 7780.80 281 omeG;omeC;rG;omeU;fluA;fluA;rA;rU;omeG;rA;omeG;fluCSup-3′ 160 5′-HO;rG;omeC;omeU;omeC;rA;fluU;fluU;omeU;fluA;fluC;rC;fluA; 7780.80 282 rG;fluC;fluG;fluU;omeA;rA;omeA;fluU;omeG;omeA;rG;omeCSup-3′ 161 5′-HO;rG;omeC;rU;fluC;rA;rU;omeU;fluU;rA;omeC;omeC;rA;omeG; 7784.93 283 omeC;fluG;rU;rA;omeA;rA;omeU;omeG;fluA;omeG;rCSup-3′ 162 5′-HO;rG;rC;rU;omeComeA;rU;omeU;omeU;omeA;fluC;omeC;omeA; 7788.85 284 fluG;rC;fluG;omeU;omeA;rA;rA;fluU;fluG;fluA;rG;omeCSup-3′ 163 5′-HO;rG;omeC;rU;omeC;omeA;fluU;fluU;omeU;omeA;fluC;omeC; 7790.86 285 fluA;omeG;rC;fluG;omeU;fluA;rA;rA;omeU;rG;rA;omeG;fluCSup-3′ 164 5′-HO;rG;omeC;fluU;omeC;fluA;omeU;omeU;fluU;fluA;rC;omeC;omeA; 7792.84 286 rG;fluC;rG;omeU;fluA;omeA;fluA;rU;omeG;omeA;rG;fluCSup-3′ 165 5′-HO;rG;omeC;omeU;omeC;fluA;omeU;omeU;omeU;rA;omeC;fluC; 7794.81 287 rA;omeG;fluC;rG;fluU;omeA;omeA;fluA;fluU;fluG;fluA;fluG;rCSup-3′ 166 5′-HO;rG;omeC;fluC;fluU;omeC;rA;fluU;omeU;fluU;fluA;fluC;omeC; 7794.85 288 omeA;omeG;fluC;omeG;rU;omeA;rA;fluA;fluU;omeG;omeA;rG; found omeCSup-3′ 167 5′-HO;rG;omeC;omeU;fluC;fluA;omeU;fluU;omeU;omeA;fluC;fluC; 7796.79 289 omeA;fluG;rC;omeG;rU;fluA;omeA;omeA;fluU;rG;fluA;fluG;omeCSup-3′ 168 5′-HO;rG;omeC;omeU;rC;fluA;omeU;omeU;rU;omeA;omeC;omeC; 7796.93 290 rA;rG;rC;rG;fluU;rA;rA;rA;omeU;omeG;omeA;fluG;omeCSup-3′ 169 5′-HO;rG;omeC;omeU;fluC;fluA;rU;omeU;omeU;omeA;omeC;rC; 7800.88 291 rA;rG;rC;omeG;omeU;omeA;rA;omeA;fluU;fluG;omeA;rG;fluCSup-3′ 170 5′-HO;rG;rC;omeU;fluC;rA;omeU;omeU;omeU;omeA;fluC;fluC;rA; Calculated 292 rG;omeC;fluG;omeU;omeA;omeA;rA;fluU;rG;omeA;fluG;omeCSup-3′ 7802.90 171 5′-HO;rG;omeC;omeU;rC;omeA;fluU;fluU;fluU;omeA;fluC;omeC;fluA; 7802.82 293 rG;rC;omeG;omeU;rA;omeA;omeA;rU;omeG;fluA;omeG;rCSup-3′ 172 5′-HO;rG;fluC;fluU;rC;omeA;fluU;omeU;omeU;rA;fluC;fluC;rA;omeG; 7802.86 294 omeC;omeG;rU;fluA;omeA;omeA;omeU;omeG;omeA;rG;rCSup-3′ 173 5′-HO;rG;omeC;rU;omeC;fluA;fluU;fluU;omeU;rA;fluC;fluC;omeA;omeG; 7808.86 295 fluC;fluG;omeU;omeA;fluA;omeA;omeU;omeG;rA;fluG;omeCSup-3′ 174 5′-HO;rG;omeC;rU;omeC;rA;omeU;omeU;rU;rA;omeC;rC;omeA;rG; 7809.00 296 omeC;fluG;rU;omeA;fluA;omeA;omeU;rG;rA;omeG;omeCSup-3′ 175 5′-HO;rG;rC;rU;fluC;omeA;omeU;rU;rU;omeA;omeC;rC;omeA;omeG; 7810.94 297 omeC;rG;rU;fluA;omeA;omeA;fluU;omeG;rA;omeG;omeCSup-3′ 176 5′-HO;rG;omeC;omeU;rC;omeA;omeU;omeU;rU;omeA;omeC;rC;omeA; 7812.88 298 omeG;omeC;fluG;fluU;rA;rA;rA;omeU;omeG;fluA;fluG;rCSup-3′ 177 5′-HO;rG;fluC;omeU;omeC;fluA;rU;rU;omeU;omeA;rC;omeC;rA;omeG; 7814.93 299 omeC;omeG;omeU;rA;fluA;fluA;omeU;rG;omeA;fluG;omeCSup-3′ 178 5′-HO;rG;omeC;omeU;omeC;fluA;rU;fluU;omeU;omeA;omeC;omeC; 7816.94 300 omeA;omeG;omeC;rG;fluU;fluA;rA;fluA;rU;omeG;omeA;rG;fluCSup-3′ 179 5′-HO;rG;dC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;omeC; 7686.65 301 rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 180 5′-HO;rG;omeC;dU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7700.68 302 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 181 5′-HO;rG;omeC;rU;dC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7700.68 303 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 182 5′-HO;rG;omeC;rU;rC;rA;dU;rU;rU;rA;fluC;rC;rA;fluG; 7698.68 304 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 183 5′-HO;rG;omeC;rU;rC;rA;fluU;dU;rU;rA;fluC;rC;rA;fluG; 7700.68 305 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 184 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;dU;rA;fluC;rC;rA;fluG; 7700.68 306 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 185 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;dC;rC;rA;fluG; 7698.68 307 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 186 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;dC;rA;fluG; 7700.68 308 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 187 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;dA;fluG; 7700.68 309 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 188 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;dG; 7698.68 310 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 189 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7686.65 311 dC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 190 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7700.68 312 omeC;dG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 191 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7698.68 313 omeC;rG;dU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 192 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7686.65 314 omeC;rG;fluU;dA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 193 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7698.68 315 omeC;rG;fluU;omeA;dA;omeA;rU;rG;rA;rG;omeCSup-3′ 194 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7686.65 316 omeC;rG;fluU;omeA;fluA;dA;rU;rG;rA;rG;omeCSup-3′ 195 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7700.68 317 omeC;rG;fluU;omeA;fluA;omeA;dU;rG;rA;rG;omeCSup-3′ 196 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7700.68 318 omeC;rG;fluU;omeA;fluA;omeA;rU;dG;rA;rG;omeCSup-3′ 197 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;omeC;rG; 7700.68 319 fluU;omeA;fluA;omeA;rU;rG;dA;rG;omeCSup-3′ 198 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7700.68 320 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;dG;omeCSup-3′ 199 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7686.65 321 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;dCSup-3′ 200 5′-HO;rG;faraC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7704.64 322 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 201 5′-HO;rG;omeC;faraU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7718.67 323 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 202 5′-HO;rG;omeC;rU;faraC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7718.67 324 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 203 5′-HO;rG;omeC;rU;rC;rA;faraU;rU;rU;rA;fluC;rC;rA;fluG; 7716.67 325 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 204 5′-HO;rG;omeC;rU;rC;rA;fluU;faraU;rU;rA;fluC;rC;rA;fluG; 7718.67 326 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 205 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;faraU;rA;fluC;rC;rA;fluG; 7718.67 327 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 206 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;faraC;rC;rA;fluG; 7716.68 328 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 207 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;faraC;rA;fluG; 7718.67 329 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 208 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7704.64 330 faraC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 209 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7716.67 331 omeC;rG;faraU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 210 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7718.67 332 omeC;rG;fluU;omeA;fluA;omeA;faraU;rG;rA;rG;omeCSup-3′ 211 5′-HO;rG;dC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7728.73 333 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 212 5′-HO;rG;rC;dU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7714.70 334 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 213 5′-HO;rG;rC;omeU;dC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7728.73 335 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 214 5′-HO;rG;rC;omeU;rC;fluA;dU;omeU;rU;rA;rC;omeC;rA;fluG; 7728.73 336 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 215 5′-HO;rG;rC;omeU;rC;fluA;rU;dU;rU;rA;rC;omeC;rA;fluG; 7714.70 337 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 216 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;dU;rA;rC;omeC;rA;fluG; 7728.73 338 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 217 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;dC;omeC;rA;fluG; 7728.73 339 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 218 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;dC;rA;fluG; 7714.70 340 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 219 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;dA;fluG; 7728.73 341 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 220 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;dG; 7726.74 342 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 221 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7714.70 343 dC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 222 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7728.73 344 omeC;dG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 223 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7714.70 345 omeC;rG;dU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 224 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7714.70 346 omeC;rG;omeU;dA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 225 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7726.74 347 omeC;rG;omeU;omeA;dA;fluA;rU;rG;omeA;rG;fluCSup-3′ 226 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7726.74 348 omeC;rG;omeU;omeA;fluA;dA;rU;rG;omeA;rG;fluCSup-3′ 227 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7728.73 349 omeC;rG;omeU;omeA;fluA;fluA;dU;rG;omeA;rG;fluCSup-3′ 228 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7728.73 350 omeC;rG;omeU;omeA;fluA;fluA;rU;dG;omeA;rG;fluCSup-3′ 229 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7714.70 351 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;dA;rG;fluCSup-3′ 230 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7728.73 352 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;dG;fluCSup-3′ 231 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7726.74 353 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;dCSup-3′ 232 5′-HO;rG;faraC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7746.72 354 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 233 5′-HO;rG;rC;faraU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7732.69 355 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 234 5′-HO;rG;rC;omeU;faraC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7746.72 356 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 235 5′-HO;rG;rC;omeU;rC;fluA;faraU;omeU;rU;rA;rC;omeC;rA;fluG; 7746.72 357 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 236 5′-HO;rG;rC;omeU;rC;fluA;rU;faraU;rU;rA;rC;omeC;rA;fluG; 7732.69 358 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 237 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;faraU;rA;rC;omeC;rA;fluG; 7746.72 359 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 238 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;faraC;omeC;rA;fluG; 7746.72 360 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 239 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;faraC;rA;fluG; 7732.69 361 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 240 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;faraA;fluG; 7746.72 362 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 241 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7732.69 363 faraC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 242 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7732.69 364 omeC;rG;faraU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 243 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7746.72 365 omeC;rG;omeU;omeA;fluA;fluA;faraU;rG;omeA;rG;fluCSup-3′ 244 5′-HO;rG;omeC;fluU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7704.64 366 omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 245 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;fluC;rA;fluG; 7704.64 367 omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 246 5′-HO;rG;omeC;fluU;rC;rA;fluU;rU;rU;rA;fluC;fluC;rA;fluG; 7706.63 368 omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 247 5′-HO;rG;omeC;rU;rC;rA;fluU;fluU;rU;rA;fluC;rC;rA;fluG; 7704.64 369 omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 248 5′-HO;rG;omeC;fluU;rC;rA;fluU;fluU;rU;rA;fluC;rC;rA;fluG; 7706.63 370 omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 249 5′-HO;rG;omeC;rU;rC;rA;fluU;fluU;rU;rA;fluC;fluC;rA;fluG; 7706.63 371 omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 250 5′-HO;rG;omeC;fluU;rC;rA;fluU;fluU;rU;rA;fluC;fluC;rA;fluG; 7708.62 372 omeC;rG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 251 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;omeC; 7716.67 373 omeG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 252 5′-HO;rG;omeC;fluU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7718.67 374 omeC;omeG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 253 5′-HO;rG;lna5meC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7728.69 375 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 254 5′-HO;rG;omeC;lna5MeU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7742.71 376 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 255 5′-HO;rG;omeC;rU;rC;lnaA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7728.69 377 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 256 5′-HO;rG;omeC;rU;rC;rA;lna5MeU;rU;rU;rA;fluC;rC;rA;fluG; 7740.72 378 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 257 5′-HO;rG;omeC;rU;rC;rA;fluU;lna5MeU;rU;rA;fluC;rC;rA;fluG; 7742.71 379 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 258 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;lna5MeU;rA;fluC;rC;rA;fluG; 7742.71 380 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 259 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;lnaA;fluC;rC;rA;fluG; 7728.69 381 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 260 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;lna5meC;rC;rA;fluG; 7740.72 382 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 261 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;lna5meC;rA;fluG; 7742.71 383 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 262 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;lnaA;fluG; 7728.69 384 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 263 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;lnaG; 7726.69 385 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 264 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7728.69 386 lna5meC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 265 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7728.69 387 omeC;lnaG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 266 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7740.72 388 omeC;rG;lna5MeU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 267 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7714.66 389 omeC;rG;fluU;lnaA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 268 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7726.69 390 omeC;rG;fluU;omeA;lnaA;omeA;rU;rG;rA;rG;omeCSup-3′ 269 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7714.66 391 omeC;rG;fluU;omeA;fluA;lnaA;rU;rG;rA;rG;omeCSup-3′ 270 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7742.71 392 omeC;rG;fluU;omeA;fluA;omeA;lna5MeU;rG;rA;rG;omeCSup-3′ 271 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7728.69 393 omeC;rG;fluU;omeA;fluA;omeA;rU;lnaG;rA;rG;omeCSup-3′ 272 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7728.69 394 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;lnaA;rG;omeCSup-3′ 273 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7728.69 395 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;lnaG;omeCSup-3′ 274 5′-HO;rG;25C;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7702.65 396 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 275 5′-HO;rG;omeC;25U;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7716.67 397 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 276 5′-HO;rG;omeC;rU;25C;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7716.68 398 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 277 5′-HO;rG;omeC;rU;rC;25A;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7716.67 399 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 278 5′-HO;rG;omeC;rU;rC;rA;25U;rU;rU;rA;fluC;rC;rA;fluG;omeC; 7714.68 400 rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 279 5′-HO;rG;omeC;rU;rC;rA;fluU;25U;rU;rA;fluC;rC;rA;fluG; 7716.67 401 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 280 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;25U;rA;fluC;rC;rA;fluG; 7716.67 402 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 281 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;25A;fluC;rC;rA;fluG; 7716.67 403 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 282 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;25C;rC;rA;fluG;omeC; 7714.69 404 rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 283 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;25C;rA;fluG; 7716.68 405 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 284 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;25A;fluG; 7716.67 406 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 285 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;25G;omeC; 7714.68 407 rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 286 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;25C; 7702.65 408 rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 287 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7714.68 409 omeC;rG;25U;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 288 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7702.65 410 omeC;rG;fluU;25A;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 289 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7714.68 411 omeC;rG;fluU;omeA;25A;omeA;rU;rG;rA;rG;omeCSup-3′ 290 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7702.65 412 omeC;rG;fluU;omeA;fluA;25A;rU;rG;rA;rG;omeCSup-3′ 291 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7716.67 413 omeC;rG;fluU;omeA;fluA;omeA;25U;rG;rA;rG;omeCSup-3′ 292 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7716.67 414 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;25A;rG;omeCSup-3′ 293 5′-HO;rG;lna5meC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7770.77 415 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 294 5′-HO;rG;rC;lna5MeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7756.74 416 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 295 5′-HO;rG;rC;omeU;lna5meC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7770.77 417 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 296 5′-HO;rG;rC;omeU;rC;lnaA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7754.75 418 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 297 5′-HO;rG;rC;omeU;rC;fluA;lna5MeU;omeU;rU;rA;rC;omeC;rA;fluG; 7770.77 419 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 298 5′-HO;rG;rC;omeU;rC;fluA;rU;lna5MeU;rU;rA;rC;omeC;rA;fluG; 7756.74 420 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 299 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;lna5MeU;rA;rC;omeC;rA;fluG; 7770.77 421 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 300 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;lnaA;rC;omeC;rA;fluG; 7756.74 422 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 301 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;lna5meC;omeC;rA;fluG; 7770.77 423 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 302 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;lna5meC;rA;fluG; 7756.74 424 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 303 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;lnaA;fluG; 7756.74 425 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 304 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;lnaG; 7754.75 426 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 305 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7756.74 427 omeC;lnaG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 306 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7756.74 428 omeC;rG;lna5MeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 307 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7742.71 429 omeC;rG;omeU;lnaA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 308 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7754.75 430 omeC;rG;omeU;omeA;lnaA;fluA;rU;rG;omeA;rG;fluCSup-3′ 309 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7754.75 431 omeC;rG;omeU;omeA;fluA;lnaA;rU;rG;omeA;rG;fluCSup-3′ 310 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7770.77 432 omeC;rG;omeU;omeA;fluA;fluA;lna5MeU;rG;omeA;rG;fluCSup-3′ 311 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7756.74 433 omeC;rG;omeU;omeA;fluA;fluA;rU;lnaG;omeA;rG;fluCSup-3′ 312 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7742.71 434 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;lnaA;rG;fluCSup-3′ 313 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7756.74 435 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;lnaG;fluCSup-3′ 314 5′-HO;rG;25C;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7744.73 436 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 315 5′-HO;rG;rC;25U;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7730.70 437 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 316 5′-HO;rG;rC;omeU;25C;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7744.73 438 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 317 5′-HO;rG;rC;omeU;rC;25A;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7742.74 439 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 318 5′-HO;rG;rC;omeU;rC;fluA;25U;omeU;rU;rA;rC;omeC;rA;fluG; 7744.73 440 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 319 5′-HO;rG;rC;omeU;rC;fluA;rU;25U;rU;rA;rC;omeC;rA;fluG; 7730.70 441 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 320 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;25U;rA;rC;omeC;rA;fluG; 7744.73 442 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 321 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;25A;rC;omeC;rA;fluG; 7744.73 443 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 322 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;25C;omeC;rA;fluG; 7744.73 444 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 323 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;25C;rA;fluG; 7730.71 445 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 324 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;25A;fluG; 7744.73 446 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 325 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;25G; 7742.74 447 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 326 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7730.71 448 25C;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 327 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7744.73 449 omeC;25G;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 328 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7730.70 450 omeC;rG;25U;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 329 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7730.70 451 omeC;rG;omeU;25A;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 330 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7742.74 452 omeC;rG;omeU;omeA;25A;fluA;rU;rG;omeA;rG;fluCSup-3′ 331 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7742.74 453 omeC;rG;omeU;omeA;fluA;25A;rU;rG;omeA;rG;fluCSup-3′ 332 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7744.73 454 omeC;rG;omeU;omeA;fluA;fluA;25U;rG;omeA;rG;fluCSup-3′ 333 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7730.70 455 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;25A;rG;fluCSup-3′ 334 5′-HO;rG;rC;omeU;rC;fluA;fluU;omeU;rU;rA;rC;omeC;rA;fluG; 7756.76 456 omeC;rG;omeU;omeA;rA;fluA;rU;rG;omeA;rG;omeCSup-3′ 335 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;fluC;omeC;rA;fluG; 7756.76 457 omeC;rG;omeU;omeA;rA;fluA;rU;rG;omeA;rG;omeCSup-3′ 336 5′-HO;rG;rC;omeU;rC;fluA;fluU;omeU;rU;rA;fluC;omeC;rA;fluG; 7758.75 458 omeC;rG;omeU;omeA;rA;fluA;rU;rG;omeA;rG;omeCSup-3′ 337 5′-HO;rG;omeC;rU;rC;r2AP;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7716.68 2 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 338 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;r2AP;fluG; 7716.68 3 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 339 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7702.65 4 omeC;rG;fluU;r2AP;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 340 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7714.68 5 omeC;rG;fluU;omeA;r2AP;omeA;rU;rG;rA;rG;omeCSup-3′ 341 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7702.65 6 omeC;rG;fluU;omeA;fluA;r2AP;rU;rG;rA;rG;omeCSup-3′ 342 5′-HO;rG;omeC;rU;rC;moeA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7774.75 459 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 343 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;moeA;fluC;rC;rA;fluG; 7774.75 460 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 344 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;moeA;fluG; 7774.75 461 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 345 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;moeG; 7772.76 462 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 346 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7774.75 463 omeC;moeG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 347 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;omeC; 7760.73 464 rG;fluU;moeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 348 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7772.76 465 omeC;rG;fluU;omeA;moeA;omeA;rU;rG;rA;rG;omeCSup-3′ 349 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7760.73 466 omeC;rG;fluU;omeA;fluA;moeA;rU;rG;rA;rG;omeCSup-3′ 350 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7774.75 467 omeC;rG;fluU;omeA;fluA;omeA;rU;moeG;rA;rG;omeCSup-3′ 351 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7774.75 468 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;moeA;rG;omeCSup-3′ 352 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7774.75 469 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;moeG;omeCSup-3′ 353 5′-HO;rG;rC;omeU;rC;r2AP;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7742.74 7 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 354 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;r2AP;rC;omeC;rA;fluG; 7744.73 8 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 355 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;r2AP;fluG; 7744.73 9 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 356 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7730.70 10 omeC;rG;omeU;r2AP;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 357 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7742.74 11 omeC;rG;omeU;omeA;r2AP;fluA;rU;rG;omeA;rG;fluCSup-3′ 358 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7742.74 12 omeC;rG;omeU;omeA;fluA;r2AP;rU;rG;omeA;rG;fluCSup-3′ 359 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7730.70 13 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;r2AP;rG;fluCSup-3′ 360 5′-HO;rG;rC;omeU;rC;moeA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7800.82 470 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 361 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;moeA;rC;omeC;rA;fluG; 7802.81 471 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 362 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;moeA;fluG; 7802.81 472 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 363 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;moeG; 7800.82 473 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 364 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7802.81 474 omeC;moeG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 365 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7788.78 475 omeC;rG;omeU;moeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 366 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7800.82 476 omeC;rG;omeU;omeA;moeA;fluA;rU;rG;omeA;rG;fluCSup-3′ 367 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7800.82 477 omeC;rG;omeU;omeA;fluA;moeA;rU;rG;omeA;rG;fluCSup-3′ 368 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7802.81 478 omeC;rG;omeU;omeA;fluA;fluA;rU;moeG;omeA;rG;fluCSup-3′ 369 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7788.78 479 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;moeA;rG;fluCSup-3′ 370 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7802.81 480 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;moeG;fluCSup-3′ 371 5′-HO;rG;omeC;fluU;rC;rA;fluU;fluU;rU;rA;fluC;rC;rA;fluG; 7720.66 481 omeC;omeG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 372 5′-HO;rG;omeC;rU;rC;rA;fluU;fluU;rU;rA;fluC;fluC;rA;fluG; 7720.66 482 omeC;omeG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 373 5′-HO;rG;omeC;fluU;rC;rA;fluU;fluU;rU;rA;fluC;fluC;rA;fluG; 7722.65 483 omeC;omeG;fluU;omeA;rA;fluA;rU;rG;rA;rG;omeCSup-3′ 374 5′-HO;rG;moe5meC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7774.75 484 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 375 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG;moe5meC 7774.75 485 rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′; 376 5′-HO;rG;omeC;rU;rC;rA;moe-5-MeU;rU;rU;rA;fluC;rC;rA;fluG; 7786.79 486 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 377 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;moe5meC;rC;rA;fluG; 7786.79 487 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 378 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7786.79 488 omeC;rG;moe5MeU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 379 5′-HO;rG;omeC;moe5MeU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7788.78 489 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 380 5′-HO;rG;omeC;rU;moe5meC;rA;fluU;rU;rU;rA;fluC;rC;rA; 7788.78 490 fluG;omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 381 5′-HO;rG;omeC;rU;rC;rA;fluU;moe5MeU;rU;rA;fluC;rC;rA;fluG; 7788.78 491 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 382 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;moe5MeU;rA;fluC;rC;rA;fluG; 7788.78 492 omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 383 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;moe5meC;rA; 7788.78 493 fluG;omeC;rG;fluU;omeA;fluA;omeA;rU;rG;rA;rG;omeCSup-3′ 384 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rU;rA;fluC;rC;rA;fluG; 7788.78 494 omeC;rG;fluU;omeA;fluA;omeA;moe5MeU;rG;rA;rG;omeCSup-3′ 385 5′-HO;rG;rC;moe5MeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7802.81 495 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 386 5′-HO;rG;rC;omeU;rC;fluA;rU;moe5MeU;rU;rA;rC;omeC;rA;fluG; 7802.81 496 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 387 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;moe5meC;rA; 7802.81 497 fluG;omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 388 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7802.81 498 moe5meC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 389 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7802.81 499 omeC;rG;moe5MeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 390 5′-HO;rG;moe5meC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7816.83 500 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 391 5′-HO;rG;rC;omeU;moe5meC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7816.83 501 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 392 5′-HO;rG;rC;omeU;rC;fluA;moe5MeU;omeU;rU;rA;rC;omeC;rA;fluG; 7816.83 502 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 393 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;moe5MeU;rA;rC;omeC;rA;fluG; 7816.83 503 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 394 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;moe5meC;omeC;rA;fluG; 7816.83 504 omeC;rG;omeU;omeA;fluA;fluA;rU;rG;omeA;rG;fluCSup-3′ 395 5′-HO;rG;rC;omeU;rC;fluA;rU;omeU;rU;rA;rC;omeC;rA;fluG; 7816.83 505 omeC;rG;omeU;omeA;fluA;fluA;moe5MeU;rG;omeA;rG;fluCSup-3′ 396 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7797.00 506 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 397 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.16 507 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 398 5′-HO;rG;omeC;rU;rC;rI;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7797.99 508 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 399 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rI;fluC;rC;rAs;fluG; 7797.99 509 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 400 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rIs;fluG; 7797.99 510 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 401 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7783.96 511 omeC;rG;fluU;rI;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 402 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7796.00 512 omeC;rG;fluU;omeA;rI;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 403 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7783.96 513 omeC;rG;fluU;omeA;fluA;rI;rUs;rG;rAs;rGs;omeCSup-3′ 404 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7797.99 514 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rIs;rGs;omeCSup-3′ 405 5′-HO;rG;d5propC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7805.03 14 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 406 5′-HO;rG;omeC;d5propU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7819.05 15 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 407 5′-HO;rG;omeC;rU;d5propC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7819.05 16 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 408 5′-HO;rG;omeC;rU;rC;rA;d5propU;rU;rUs;rA;fluC;rC;rAs;fluG; 7817.06 17 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 409 5′-HO;rG;omeC;rU;rC;rA;fluU;d5propU;rUs;rA;fluC;rC;rAs;fluG; 7819.05 18 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 410 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;d5propUs;rA;fluC;rC;rAs;fluG; 7819.05 19 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 411 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;d5propC;rC;rAs;fluG; 7817.06 20 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 412 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;d5propC;rAs;fluG; 7819.05 21 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 413 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7805.03 22 d5propC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 414 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7817.06 23 rG;d5propU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 415 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7819.05 24 rG;fluU;omeA;fluA;omeA;d5propUs;rG;rAs;rGs;omeCSup-3′ 416 5′-HO;rG;omeC;rU;rC;r7dz8azA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7797.01 25 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 417 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;r7dz8azA;fluC;rC;rAs;fluG; 7797.01 26 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 418 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;r7dz8azAs;fluG; 7797.00 27 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 419 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7782.98 28 rG;fluU;r7dz8azA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 420 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7795.01 29 rG;fluU;omeA;r7dz8azA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 421 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7782.98 30 rG;fluU;omeA;fluA;r7dz8azA;rUs;rG;rAs;rGs;omeCSup-3′ 422 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7797.00 31 rG;fluU;omeA;fluA;omeA;rUs;rG;r7dz8azAs;rGs;omeCSup-3′ 423 5′-HO;rG;omeC;rU;rC;d2aminoA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7796.02 32 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 424 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;d2aminoA;fluC;rC;rAs;fluG; 7796.02 33 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 425 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;d2aminoAs;fluG; 7796.02 34 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 426 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7781.99 35 fluU;d2aminoA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 427 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7794.03 36 fluU;omeA;d2aminoA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 428 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7781.99 37 fluU;omeA;fluA;d2aminoA;rUs;rG;rAs;rGs;omeCSup-3′ 429 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7796.02 38 fluU;omeA;fluA;omeA;rUs;rG;d2aminoAs;rGs;omeCSup-3′ 430 5′-HO;rG;omeC;rU;rC;fara8azA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7799.98 39 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 431 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;fara8azA;fluC;rC;rAs;fluG; 7799.98 40 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 432 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;fara8azAs;fluG;omeC; 7799.98 41 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 433 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7785.96 42 fluU;fara8azA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 434 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7797.99 43 fluU;omeA;fara8azA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 435 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7785.96 44 fluU;omeA;fluA;fara8azA;rUs;rG;rAs;rGs;omeCSup-3′ 436 5′-HO;rG;rC;rUs;fluC;rI;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC; 7860.12 515 omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 437 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;rI;omeC;omeC;rAs;omeG; 7872.15 516 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 438 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rIs;omeG; 7874.14 517 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 439 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7874.14 518 omeC;omeG;omeU;rI;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 440 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7872.15 519 omeC;omeG;omeU;rA;rI;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 441 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7872.15 520 omeC;omeG;omeU;rA;fluA;rI;omeUs;fluG;omeAs;rGs;fluCSup-3′ 442 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7860.12 521 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;rIs;rGs;fluCSup-3′ 443 5′-HO;rG;d5propC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7895.21 45 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 444 5′-HO;rG;rC;d5propUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7895.20 46 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 445 5′-HO;rG;rC;rUs;d5propC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7893.22 47 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 446 5′-HO;rG;rC;rUs;fluC;omeA;d5propU;rU;rUs;fluA;omeC;omeC;rAs; 7893.22 48 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 447 5′-HO;rG;rC;rUs;fluC;omeA;fluU;d5propU;rUs;fluA;omeC;omeC;rAs; 7895.21 49 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 448 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;d5propUs;fluA;omeC;omeC;rAs; 7895.20 50 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 449 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;d5propC;omeC;rAs;omeG; 7881.18 51 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 450 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;d5propC;rAs;omeG; 7881.18 52 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 451 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7881.18 53 d5propC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 452 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7881.18 54 omeC;omeG;d5propU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 453 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7881.18 55 omeC;omeG;omeU;rA;fluA;fluA;d5propUs;fluG;omeAs;rGs;fluCSup-3′ 454 5′-HO;rG;rC;rUs;fluC;r7dz8azA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7859.13 56 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 455 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;r7dz8azA;omeC;omeC;rAs; 7871.17 57 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 456 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;r7dz8azAs; 7873.16 58 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 457 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.16 59 omeC;omeG;omeU;r7dz8azA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 458 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7871.17 60 omeC;omeG;omeU;rA;r7dz8azA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 459 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7871.17 61 omeC;omeG;omeU;rA;fluA;r7dz8azA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 460 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7859.13 62 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;r7dz8azAs;rGs;fluCSup-3′ 461 5′-HO;rG;rC;rUs;fluC;d2aminoA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7858.15 63 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 462 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;d2aminoA;omeC;omeC;rAs; 7870.18 64 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 463 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;d2aminoAs; 7872.17 65 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 464 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7872.18 66 omeC;omeG;omeU;d2aminoA;fluA;fluA;omeUs;fluG;omeAs;rGs; fluCSup-3′ 465 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7870.18 67 omeC;omeG;omeU;rA;d2aminoA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 466 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7870.18 68 omeC;omeG;omeU;rA;fluA;d2aminoA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 467 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7858.14 69 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;d2aminoAs;rGs;fluCSup-3′ 468 5′-HO;rG;rC;rUs;fluC;fara8azA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7862.11 70 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 469 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fara8azA;omeC;omeC;rAs; 7874.15 71 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 470 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;fara8azAs; 7876.13 72 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 471 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7876.14 73 omeC;omeG;omeU;fara8azA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 472 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7874.15 74 omeC;omeG;omeU;rA;fara8azA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 473 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7874.15 75 omeC;omeG;omeU;rA;fluA;fara8azA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 474 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7862.11 76 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;fara8azAs;rGs;fluCSup-3′ 475 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;fluC;rAs;fluG;omeC; 7798.99 522 omeG;fluU;omeA;rA;fluA;rUs;rG;rAs;rGs;omeCSup-3′ 476 5′-HO;rG;omeC;fluU;rC;rA;fluU;rU;rUs;rA;fluC;fluC;rAs;fluG;omeC; 7800.99 523 omeG;fluU;omeA;rA;fluA;rUs;rG;rAs;rGs;omeCSup-3′ 477 5′-HO;rG;omeC;rU;rC;rA;fluU;fluU;rUs;rA;fluC;rC;rAs;fluG; 7798.99 524 omeC;omeG;fluU;omeA;rA;fluA;rUs;rG;rAs;rGs;omeCSup-3′ 478 5′-HO;rG;omeC;rU;rC;fara7dzA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7798.01 77 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 479 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;fara7dzA;fluC;rC;rAs;fluG; 7798.01 78 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 480 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;fara7dzAs;fluG;omeC; 7798.00 79 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 481 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7783.98 80 omeC;rG;fluU;fara7dzA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 482 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7796.02 81 fluU;omeA;fara7dzA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 483 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7783.98 82 omeC;rG;fluU;omeA;fluA;fara7dzA;rUs;rG;rAs;rGs;omeCSup-3′ 484 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7798.00 83 fluU;omeA;fluA;omeA;rUs;rG;fara7dzAs;rGs;omeCSup-3′ 485 5′-HO;rG;omeC;rU;rC;d7dzA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7780.02 84 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 486 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;d7dzA;fluC;rC;rAs;fluG; 7780.02 85 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 487 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;d7dzAs;fluG; 7780.01 86 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 488 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7765.99 87 omeC;rG;fluU;d7dzA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 489 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7778.03 88 omeC;rG;fluU;omeA;d7dzA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 490 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7765.99 89 omeC;rG;fluU;omeA;fluA;d7dzA;rUs;rG;rAs;rGs;omeCSup-3′ 491 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7780.01 90 fluU;omeA;fluA;omeA;rUs;rG;d7dzAs;rGs;omeCSup-3′ 492 5′-HO;rG;omeC;rU;rC;r7dzA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7796.02 91 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 493 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;r7dzA;fluC;rC;rAs;fluG; 7796.02 92 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 494 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;r7dzAs;fluG; 7796.01 93 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 495 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7781.99 94 omeC;rG;fluU;r7dzA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 496 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7794.03 95 omeC;rG;fluU;omeA;r7dzA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 497 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7781.99 96 omeC;rG;fluU;omeA;fluA;r7dzA;rUs;rG;rAs;rGs;omeCSup-3′ 498 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7796.01 97 fluU;omeA;fluA;omeA;rUs;rG;r7dzAs;rGs;omeCSup-3′ 499 5′-HO;rG;rC;rUs;fluC;fara7dzA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7860.14 98 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 500 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fara7dzA;omeC;omeC;rAs; 7872.17 99 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 501 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;fara7dzAs; 7874.16 100 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 502 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7874.16 101 omeC;omeG;omeU;fara7dzA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 503 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7872.17 102 omeC;omeG;omeU;rA;fara7dzA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 504 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7872.17 103 omeC;omeG;omeU;rA;fluA;fara7dzA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 505 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7860.13 104 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;fara7dzAs;rGs;fluCSup-3′ 506 5′-HO;rG;rC;rUs;fluC;d7dzA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7842.15 105 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 507 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;d7dzA;omeC;omeC;rAs;omeG; 7854.18 106 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 508 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;d7dzAs; 7856.17 107 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 509 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7856.17 108 omeC;omeG;omeU;d7dzA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 510 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7854.18 109 omeC;omeG;omeU;rA;d7dzA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 511 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7854.18 110 omeC;omeG;omeU;rA;fluA;d7dzA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 512 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7842.14 111 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;d7dzAs;rGs;fluCSup-3′ 513 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;r7dzA;omeC;omeC;rAs;omeG; 7870.18 112 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 514 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;fluC;omeC;rAs;omeG; 7843.11 525 omeC;omeG;omeU;rA;rA;fluA;rUs;rG;omeAs;rGs;fluCSup-3′ 515 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7855.15 526 omeC;omeG;omeU;rA;rA;fluA;rUs;rG;omeAs;rGs;fluCSup-3′ 516 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;fluC;omeC;rAs;omeG; 7845.10 527 omeC;omeG;omeU;rA;fluA;fluA;rUs;rG;omeAs;rGs;fluCSup-3′ 517 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;fluC;omeC;rAs;omeG; 7857.14 528 omeC;omeG;omeU;rA;rA;fluA;omeUs;rG;omeAs;rGs;fluCSup-3′ 518 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;fluC;omeC;rAs;omeG; 7845.10 529 omeC;omeG;omeU;rA;rA;fluA;rUs;fluG;omeAs;rGs;fluCSup-3′ 519 5′-HO;thiorG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7813.07 530 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 520 5′-HO;rG;omeC;thiorU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7813.06 531 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 521 5′-HO;rG;omeC;rU;thiorC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7813.07 532 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 522 5′-HO;rG;omeC;rU;rC;thiorA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7813.07 533 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 523 5′-HO;rG;omeC;rU;rC;rA;thiorU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7811.07 534 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 524 5′-HO;rG;omeC;rU;rC;rA;fluU;thiorU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7813.06 535 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 525 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;thiorUs;rA;fluC;rC;rAs;fluG;omeC; 7813.06 536 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 526 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;thiorA;fluC;rC;rAs;fluG;omeC; 7813.07 537 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 527 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;thiorC;rC;rAs;fluG;omeC; 7811.07 538 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 528 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;thiorC;rAs;fluG;omeC; 7813.07 539 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 529 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;thiorAs;fluG;omeC; 7813.06 540 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 530 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;thiorG;omeC; 7811.08 541 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 531 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;thiorC;rG; 7799.04 542 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 532 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7813.07 543 thiorG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 533 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7811.07 544 thiorU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 534 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7799.04 545 fluU;thiorA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 535 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7811.08 546 fluU;omeA;thiorA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 536 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7799.04 547 fluU;omeA;fluA;thiorA;rUs;rG;rAs;rGs;omeCSup-3′ 537 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7813.06 548 fluU;omeA;fluA;omeA;thiorUs;rG;rAs;rGs;omeCSup-3′ 538 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7813.07 549 fluU;omeA;fluA;omeA;rUs;thiorG;rAs;rGs;omeCSup-3′ 539 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7813.06 550 fluU;omeA;fluA;omeA;rUs;rG;thiorAs;rGs;omeCSup-3′ 540 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7813.06 551 fluU;omeA;fluA;omeA;rUs;rG;rAs;thiorGs;omeCSup-3′ 541 5′-HO;rG;cf3C;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7850.97 552 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 542 5′-HO;rG;omeC;cf3U;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7865.00 553 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 543 5′-HO;rG;omeC;rU;cf3C;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7865.00 554 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 544 5′-HO;rG;omeC;rU;rC;rA;cf3U;rU;rUs;rA;fluC;rC;rAs;fluG; 7863.01 555 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 545 5′-HO;rG;omeC;rU;rC;rA;fluU;cf3U;rUs;rA;fluC;rC;rAs;fluG; 7865.00 556 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 546 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;cf3Us;rA;fluC;rC;rAs;fluG; 7865.00 557 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 547 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;cf3C;rC;rAs;fluG; 7863.01 558 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 548 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;cf3C;rAs;fluG; 7865.00 559 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 549 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;cf3As;fluG; 7865.00 560 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 550 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7850.97 561 cf3C;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 551 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7863.01 562 omeC;rG;cf3U;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 552 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7850.97 563 omeC;rG;fluU;cf3A;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 553 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7863.01 564 omeC;rG;fluU;omeA;cf3A;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 554 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7865.00 565 omeC;rG;fluU;omeA;fluA;omeA;cf3Us;rG;rAs;rGs;omeCSup-3′ 555 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7865.00 566 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;cf3As;rGs;omeCSup-3′ 556 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG; 7865.00 567 omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;cf3Gs;omeCSup-3′ 557 5′-HO;thiorG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7889.22 568 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 558 5′-HO;rG;thiorC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7889.22 569 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 559 5′-HO;rG;rC;thiorUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7889.21 570 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 560 5′-HO;rG;rC;rUs;fluC;omeA;thiorU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7887.23 571 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 561 5′-HO;rG;rC;rUs;fluC;omeA;fluU;thiorU;rUs;fluA;omeC;omeC;rAs; 7889.22 572 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 562 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;thiorUs;fluA;omeC;omeC;rAs; 7889.21 573 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 563 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;thiorA;omeC;omeC;rAs;omeG; 7887.23 574 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 564 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;thiorC;omeC;rAs;omeG; 7875.19 575 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 565 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;thiorC;rAs;omeG; 7875.19 576 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 566 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;thiorAs; 7889.22 577 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 567 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;thiorG; 7875.19 578 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 568 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7875.19 579 thiorC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 569 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7875.19 580 omeC;thiorG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 570 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7875.19 581 omeC;omeG;thiorU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 571 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7889.22 582 omeC;omeG;omeU;thiorA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 572 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7887.23 583 omeC;omeG;omeU;rA;thiorA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 573 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7887.23 584 omeC;omeG;omeU;rA;fluA;thiorA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 574 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7875.19 585 omeC;omeG;omeU;rA;fluA;fluA;thiorUs;fluG;omeAs;rGs;fluCSup-3′ 575 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7887.23 586 omeC;omeG;omeU;rA;fluA;fluA;omeUs;thiorG;omeAs;rGs;fluCSup-3′ 576 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7875.19 587 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;thiorAs;rGs;fluCSup-3′ 577 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7889.22 588 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;thiorGs;fluCSup-3′ 578 5′-HO;rG;cf3C;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7941.16 589 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 579 5′-HO;rG;rC;cf3Us;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7941.16 590 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 580 5′-HO;rG;rC;rUs;cf3C;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7939.16 591 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 581 5′-HO;rG;rC;rUs;fluC;omeA;cf3U;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7939.16 592 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 582 5′-HO;rG;rC;rUs;fluC;omeA;fluU;cf3U;rUs;fluA;omeC;omeC;rAs;omeG; 7941.16 593 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 583 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;cf3Us;fluA;omeC;omeC;rAs;omeG; 7941.16 594 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 584 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;cf3A;omeC;omeC;rAs;omeG; 7939.16 595 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 585 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;cf3C;omeC;rAs;omeG; 7927.13 596 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 586 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;cf3C;rAs;omeG; 7927.13 597 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 587 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;cf3As;omeG; 7941.16 598 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 588 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7927.13 599 cf3C;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 589 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7927.13 600 omeC;omeG;cf3U;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 590 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7939.16 601 omeC;omeG;omeU;rA;cf3A;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 591 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7939.16 602 omeC;omeG;omeU;rA;fluA;cf3A;omeUs;fluG;omeAs;rGs;fluCSup-3′ 592 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7927.13 603 omeC;omeG;omeU;rA;fluA;fluA;cf3Us;fluG;omeAs;rGs;fluCSup-3′ 593 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7927.13 604 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;cf3As;rGs;fluCSup-3′ 594 5′-HO;4S8azG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7814.05 113 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 595 5′-HO;rG;araC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7782.98 605 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 596 5′-HO;rG;omeC;araU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7797.00 606 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 597 5′-HO;rG;omeC;rU;araC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7797.00 607 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 598 5′-HO;rG;omeC;rU;rC;araA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7797.00 608 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 599 5′-HO;rG;omeC;rU;rC;rA;araU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7795.01 609 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 600 5′-HO;rG;omeC;rU;rC;rA;fluU;araU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7797.00 610 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 601 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;araUs;rA;fluC;rC;rAs;fluG;omeC; 7797.00 611 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 602 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;araA;fluC;rC;rAs;fluG;omeC; 7797.00 612 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 603 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;araC;rC;rAs;fluG;omeC;rG; 7795.01 613 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 604 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;araC;rAs;fluG;omeC; 7797.00 614 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 605 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;araAs;fluG;omeC; 7797.00 615 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 606 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;4S8azG;omeC; 7812.06 114 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 607 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;araC;rG; 7782.98 616 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 608 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7814.05 115 4S8azG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 609 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7795.01 617 araU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 610 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7782.98 618 fluU;araA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 611 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7795.01 619 fluU;omeA;araA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 612 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7782.98 620 fluU;omeA;fluA;araA;rUs;rG;rAs;rGs;omeCSup-3′ 613 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7797.00 621 fluU;omeA;fluA;omeA;araUs;rG;rAs;rGs;omeCSup-3′ 614 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7814.05 116 fluU;omeA;fluA;omeA;rUs;4S8azG;rAs;rGs;omeCSup-3′ 615 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7797.00 622 fluU;omeA;fluA;omeA;rUs;rG;araAs;rGs;omeCSup-3′ 616 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7814.05 117 fluU;omeA;fluA;omeA;rUs;rG;rAs;4S8azGs;omeCSup-3′ 617 5′-HO;rG;omeC;rU;rC;rA;rxyloU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7795.01 623 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 618 5′-HO;rG;omeC;rU;rC;rA;fluU;rxyloU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7797.00 624 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 619 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rxyloUs;rA;fluC;rC;rAs;fluG;omeC; 7797.00 625 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 620 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;rxyloC;rC;rAs;fluG;omeC; 7795.01 626 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 621 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rxyloC;rAs;fluG;omeC; 7797.00 627 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 622 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rxyloAs;fluG;omeC; 7797.00 628 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 623 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;rxyloG;omeC; 7795.01 629 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 624 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;rxyloC;rG; 7782.98 630 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 625 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rxyloG; 7797.00 631 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 626 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7795.01 632 rxyloU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 627 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7782.98 633 fluU;rxyloA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 628 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7795.01 634 fluU;omeA;rxyloA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 629 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7782.98 635 fluU;omeA;fluA;rxyloA;rUs;rG;rAs;rGs;omeCSup-3′ 630 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7797.00 636 fluU;omeA;fluA;omeA;rxyloUs;rG;rAs;rGs;omeCSup-3′ 631 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7797.00 637 fluU;omeA;fluA;omeA;rUs;rxyloG;rAs;rGs;omeCSup-3′ 632 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7797.00 638 fluU;omeA;fluA;omeA;rUs;rG;rxyloAs;rGs;omeCSup-3′ 633 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7797.00 639 fluU;omeA;fluA;omeA;rUs;rG;rAs;rxyloGs;omeCSup-3′ 634 5′-HO;4S8azG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7890.21 118 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 635 5′-HO;rG;araC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.16 640 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 636 5′-HO;rG;rC;araUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.16 641 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 637 5′-HO;rG;rC;rUs;araC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7871.17 642 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 638 5′-HO;rG;rC;rUs;fluC;araA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7859.13 643 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 639 5′-HO;rG;rC;rUs;fluC;omeA;araU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7871.17 644 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 640 5′-HO;rG;rC;rUs;fluC;omeA;fluU;araU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.16 645 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 641 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;araUs;fluA;omeC;omeC;rAs;omeG; 7873.16 646 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 642 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;araA;omeC;omeC;rAs;omeG; 7871.17 647 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 643 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;araC;omeC;rAs;omeG; 7859.13 648 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 644 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;araC;rAs;omeG; 7859.13 649 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 645 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;araAs;omeG; 7873.16 650 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 646 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;4S8azG; 7876.18 119 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 647 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7859.13 651 araC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 648 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7876.18 120 omeC;4S8azG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 649 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7859.13 652 omeC;omeG;araU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 650 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.16 653 omeC;omeG;omeU;araA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 651 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7871.17 654 omeC;omeG;omeU;rA;araA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 652 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7871.17 655 omeC;omeG;omeU;rA;fluA;araA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 653 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7859.13 656 omeC;omeG;omeU;rA;fluA;fluA;araUs;fluG;omeAs;rGs;fluCSup-3′ 654 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7888.22 121 omeC;omeG;omeU;rA;fluA;fluA;omeUs;4S8azG;omeAs;rGs;fluCSup-3′ 655 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7859.13 657 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;araAs;rGs;fluCSup-3′ 656 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7890.20 122 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;4S8azGs;fluCSup-3′ 657 5′-HO;rG;rC;rUs;fluC;omeA;rxyloU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7871.17 658 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 658 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rxyloU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.16 659 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 659 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rxyloUs;fluA;omeC;omeC;rAs;omeG; 7873.16 660 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 660 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;rxyloC;rAs;omeG; 7859.13 661 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 661 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rxyloAs; 7873.16 662 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 662 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;rxyloG; 7859.13 663 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 663 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7859.13 664 rxyloC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 664 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7859.13 665 omeC;rxyloG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 665 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7859.13 666 omeC;omeG;rxyloU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 666 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.16 667 omeC;omeG;omeU;rxyloA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 667 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7871.17 668 omeC;omeG;omeU;rA;rxyloA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 668 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7871.17 669 omeC;omeG;omeU;rA;fluA;rxyloA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 669 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7859.13 670 omeC;omeG;omeU;rA;fluA;fluA;rxyloUs;fluG;omeAs;rGs;fluCSup-3′ 670 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7871.17 671 omeC;omeG;omeU;rA;fluA;fluA;omeUs;rxyloG;omeAs;rGs;fluCSup-3′ 671 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7859.13 672 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;rxyloAs;rGs;fluCSup-3′ 672 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.16 673 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rxyloGs;fluCSup-3′ 673 5′-HO;amiG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7796.02 674 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 674 5′-HO;rG;amiC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7781.99 675 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 675 5′-HO;rG;omeC;amiU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7796.02 676 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 676 5′-HO;rG;omeC;rU;rC;amiA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7796.02 677 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 677 5′-HO;rG;omeC;rU;rC;rA;amiU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7794.03 678 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 678 5′-HO;rG;omeC;rU;rC;rA;fluU;amiU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7796.02 679 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 679 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;amiUs;rA;fluC;rC;rAs;fluG;omeC; 7796.02 680 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 680 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;amiC;rC;rAs;fluG;omeC;rG; 7794.03 681 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 681 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;amiAs;fluG;omeC; 7796.02 682 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 682 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;amiG;omeC;rG; 7794.03 683 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 683 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;amiC;rG; 7781.99 684 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 684 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7794.03 685 amiU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 685 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7794.03 686 fluU;omeA;amiA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 686 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7781.99 687 fluU;omeA;fluA;amiA;rUs;rG;rAs;rGs;omeCSup-3′ 687 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7796.02 688 fluU;omeA;fluA;omeA;amiUs;rG;rAs;rGs;omeCSup-3′ 688 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7796.02 689 fluU;omeA;fluA;omeA;rUs;amiG;rAs;rGs;omeCSup-3′ 689 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7796.02 690 fluU;omeA;fluA;omeA;rUs;rG;amiAs;rGs;omeCSup-3′ 690 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7796.02 691 fluU;omeA;fluA;omeA;rUs;rG;rAs;amiGs;omeCSup-3′ 691 5′-HO;rG;unaC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7784.99 692 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 692 5′-HO;rG;omeC;unaU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7799.02 693 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 693 5′-HO;rG;omeC;rU;unaC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7799.02 694 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 694 5′-HO;rG;omeC;rU;rC;unaA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7799.02 695 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 695 5′-HO;rG;omeC;rU;rC;rA;unaU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7797.03 696 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 696 5′-HO;rG;omeC;rU;rC;rA;fluU;unaU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7799.02 697 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 697 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;unaUs;rA;fluC;rC;rAs;fluG;omeC; 7799.02 698 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 698 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;unaA;fluC;rC;rAs;fluG;omeC; 7799.02 699 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 699 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;unaC;rC;rAs;fluG;omeC;rG; 7797.03 700 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 700 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;unaC;rAs;fluG;omeC; 7799.02 701 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 701 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;unaAs;fluG;omeC; 7799.02 702 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 702 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;unaG;omeC;rG; 7797.03 703 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 703 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;unaC;rG; 7784.99 704 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 704 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;unaG; 7799.02 705 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 705 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7797.03 706 unaU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 706 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7784.99 707 fluU;unaA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 707 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7797.03 708 fluU;omeA;unaA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 708 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7784.99 709 fluU;omeA;fluA;unaA;rUs;rG;rAs;rGs;omeCSup-3′ 709 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7799.02 710 fluU;omeA;fluA;omeA;unaUs;rG;rAs;rGs;omeCSup-3′ 710 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7799.02 711 fluU;omeA;fluA;omeA;rUs;unaG;rAs;rGs;omeCSup-3′ 711 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7799.02 712 fluU;omeA;fluA;omeA;rUs;rG;unaAs;rGs;omeCSup-3′ 712 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7799.02 713 fluU;omeA;fluA;omeA;rUs;rG;rAs;unaGs;omeCSup-3′ 713 5′-HO;amiG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7872.17 714 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 714 5′-HO;rG;amiC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7872.17 715 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 715 5′-HO;rG;rC;amiUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7872.17 716 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 716 5′-HO;rG;rC;rUs;amiC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7870.18 717 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 717 5′-HO;rG;rC;rUs;fluC;amiA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7858.15 718 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 718 5′-HO;rG;rC;rUs;fluC;omeA;amiU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7870.18 719 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 719 5′-HO;rG;rC;rUs;fluC;omeA;fluU;amiU;rUs;fluA;omeC;omeC;rAs;omeG; 7872.17 720 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 720 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;amiUs;fluA;omeC;omeC;rAs;omeG; 7872.17 721 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 721 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;amiC;omeC;rAs;omeG; 7858.15 722 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 722 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;amiC;rAs;omeG; 7858.15 723 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 723 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;amiAs;omeG; 7872.17 724 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 724 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;amiG; 7858.15 725 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 725 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7858.15 726 amiC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 726 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7858.15 727 omeC;amiG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 727 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7858.15 728 omeC;omeG;amiU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 728 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7872.17 729 omeC;omeG;omeU;amiA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 729 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7870.18 730 omeC;omeG;omeU;rA;amiA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 730 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7870.18 731 omeC;omeG;omeU;rA;fluA;amiA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 731 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7858.15 732 omeC;omeG;omeU;rA;fluA;fluA;amiUs;fluG;omeAs;rGs;fluCSup-3′ 732 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7858.15 733 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;amiAs;rGs;fluCSup-3′ 733 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7872.17 734 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;amiGs;fluCSup-3′ 734 5′-HO;unaG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7875.17 735 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 735 5′-HO;rG;unaC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7875.17 736 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 736 5′-HO;rG;rC;unaUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7875.17 737 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 737 5′-HO;rG;rC;rUs;unaC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.18 738 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 738 5′-HO;rG;rC;rUs;fluC;unaA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7861.15 739 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 739 5′-HO;rG;rC;rUs;fluC;omeA;unaU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.18 740 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 740 5′-HO;rG;rC;rUs;fluC;omeA;fluU;unaU;rUs;fluA;omeC;omeC;rAs;omeG; 7875.17 741 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 741 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;unaUs;fluA;omeC;omeC;rAs;omeG; 7875.17 742 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 742 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;unaA;omeC;omeC;rAs;omeG; 7873.18 743 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 743 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;unaC;omeC;rAs;omeG; 7861.15 744 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 744 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;unaC;rAs;omeG; 7861.15 745 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 745 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;unaG; 7861.15 746 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 746 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7861.15 747 omeC;omeG;unaU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 747 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7875.17 748 omeC;omeG;omeU;unaA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 748 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.18 749 omeC;omeG;omeU;rA;unaA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 749 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.18 750 omeC;omeG;omeU;rA;fluA;unaA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 750 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7861.15 751 omeC;omeG;omeU;rA;fluA;fluA;unaUs;fluG;omeAs;rGs;fluCSup-3′ 751 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7873.18 752 omeC;omeG;omeU;rA;fluA;fluA;omeUs;unaG;omeAs;rGs;fluCSup-3′ 752 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7861.15 753 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;unaAs;rGs;fluCSup-3′ 753 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7875.17 754 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;unaGs;fluCSup-3′ 754 5′-HO;4alk- 7805.03 755 dG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU; omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 755 5′-HO;rG;4alk- 7791.00 756 dC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA; fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 756 5′-HO;rG;omeC;4alk- 7805.03 757 dU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA; fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 757 5′-HO;rG;omeC;rU;4alk- 7805.03 758 dC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA; omeA;rUs;rG;rAs;rGs;omeCSup-3′ 758 5′-HO;rG;omeC;rU;rC;4alk- 7805.03 759 dA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA; rUs;rG;rAs;rGs;omeCSup-3′ 759 5′-HO;rG;omeC;rU;rC;rA;4alk- 7803.04 760 dU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA; rUs;rG;rAs;rGs;omeCSup-3′ 760 5′-HO;rG;omeC;rU;rC;rA;fluU;4alk- 7805.03 761 dU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;rUs; rG;rAs;rGs;omeCSup-3′ 761 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;4alk- 7805.02 762 dUs;rA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;rUs;rG; rAs;rGs;omeCSup-3′ 762 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;4alk- 7805.03 763 dA;fluC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs; rGs;omeCSup-3′ 763 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;4alk- 7803.04 764 dC;rC;rAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs; omeCSup-3′ 764 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;4alk- 7805.02 765 dAs;fluG;omeC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 765 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;4alk- 7791.00 766 dC;rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 766 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7805.03 767 4alk-dG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 767 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7803.04 768 rG;4alk-dU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 768 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7791.00 769 fluU;4alk-dA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 769 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7803.04 770 fluU;omeA;4alk-dA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 770 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7791.00 771 fluU;omeA;fluA;4alk-dA;rUs;rG;rAs;rGs;omeCSup-3′ 771 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7805.02 772 fluU;omeA;fluA;omeA;4alk-dUs;rG;rAs;rGs;omeCSup-3′ 772 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7805.02 773 fluU;omeA;fluA;omeA;rUs;rG;4alk-dAs;rGs;omeCSup-3′ 773 5′-HO;ome4SG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7827.09 774 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 774 5′-HO;rG;ome4SC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7813.07 775 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 775 5′-HO;rG;omeC;ome4SU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7827.09 776 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 776 5′-HO;rG;omeC;rU;ome4SC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7827.09 777 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 777 5′-HO;rG;omeC;rU;rC;ome4SA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7827.09 778 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 778 5′-HO;rG;omeC;rU;rC;rA;ome4SU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7825.10 779 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 779 5′-HO;rG;omeC;rU;rC;rA;fluU;ome4SU;rUs;rA;fluC;rC;rAs;fluG;omeC; 7827.09 780 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 780 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;ome4SUs;rA;fluC;rC;rAs;fluG;omeC; 7827.09 781 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 781 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;ome4SC;rC;rAs;fluG;omeC; 7825.10 782 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 782 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;ome4SG;omeC; 7825.10 783 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 783 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;ome4SC; 7813.07 784 rG;fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 784 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;ome4SG; 7827.09 785 fluU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 785 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7825.10 786 ome4SU;omeA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 786 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7813.07 787 fluU;ome4SA;fluA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 787 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7825.10 788 fluU;omeA;ome4SA;omeA;rUs;rG;rAs;rGs;omeCSup-3′ 788 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7813.07 789 fluU;omeA;fluA;ome4SA;rUs;rG;rAs;rGs;omeCSup-3′ 789 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7827.09 790 fluU;omeA;fluA;omeA;ome4SUs;rG;rAs;rGs;omeCSup-3′ 790 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7827.09 791 fluU;omeA;fluA;omeA;rUs;ome4SG;rAs;rGs;omeCSup-3′ 791 5′-HO;rG;omeC;rU;rC;rA;fluU;rU;rUs;rA;fluC;rC;rAs;fluG;omeC;rG; 7827.09 792 fluU;omeA;fluA;omeA;rUs;rG;rAs;ome4SGs;omeCSup-3′ 792 5′-HO;4alk- 7881.18 793 dG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC; omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 793 5′-HO;rG;4alk- 7881.18 794 dC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC; omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 794 5′-HO;rG;rC;4alk- 7881.18 795 dUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC; omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 795 5′-HO;rG;rC;rUs;4alk- 7879.19 844 dC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG; omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 796 5′-HO;rG;rC;rUs;fluC;4alk- 7867.16 796 dA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU; rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 797 5′-HO;rG;rC;rUs;fluC;omeA;4alk- 7879.19 797 dU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA; fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 798 5′-HO;rG;rC;rUs;fluC;omeA;fluU;4alk- 7881.18 798 dU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA; fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 799 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;4alk- 7881.18 799 dUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA;fluA; omeUs;fluG;omeAs;rGs;fluCSup-3′ 800 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;4alk- 7879.19 800 dA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA;fluA;ome Us;fluG;omeAs;rGs;fluCSup-3′ 801 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;4alk- 7881.18 801 dAs;omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs; rGs;fluCSup-3′ 802 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7867.16 802 omeC;4alk-dG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 803 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7867.15 803 omeC;omeG;4alk-dU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 804 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7881.18 804 omeC;omeG;omeU;4alk-dA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 805 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7879.19 805 omeC;omeG;omeU;rA;4alk-dA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 806 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7879.19 806 omeC;omeG;omeU;rA;fluA;4alk-dA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 807 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7867.15 807 omeC;omeG;omeU;rA;fluA;fluA;4alk-dUs;fluG;omeAs;rGs;fluCSup-3′ 808 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7879.19 808 omeC;omeG;omeU;rA;fluA;fluA;omeUs;4alk-dG;omeAs;rGs;fluCSup-3′ 809 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7867.15 809 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;4alk-dAs;rGs;fluCSup-3′ 810 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7881.18 810 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;4alk-dGs;fluCSup-3′ 811 5′-HO;ome4SG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7903.25 811 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 812 5′-HO;rG;ome4SC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7903.25 812 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 813 5′-HO;rG;rC;ome4SUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7903.24 813 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 814 5′-HO;rG;rC;rUs;ome4SC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs; 7901.26 814 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 815 5′-HO;rG;rC;rUs;fluC;omeA;ome4SU;rU;rUs;fluA;omeC;omeC;rAs; 7901.25 815 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 816 5′-HO;rG;rC;rUs;fluC;omeA;fluU;ome4SU;rUs;fluA;omeC;omeC;rAs; 7903.25 816 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 817 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;ome4SUs;fluA;omeC;omeC;rAs; 7903.24 817 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 818 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;ome4SC;omeC;rAs;omeG; 7889.22 818 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 819 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;ome4SAs; 7903.24 819 omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 820 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;ome4SG; 7889.22 820 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 821 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7889.22 821 ome4SC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 822 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7889.22 822 omeC;ome4SG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 823 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7889.22 823 omeC;omeG;ome4SU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 824 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7903.25 824 omeC;omeG;omeU;ome4SA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 825 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7901.26 825 omeC;omeG;omeU;rA;ome4SA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ 826 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7889.21 826 omeC;omeG;omeU;rA;fluA;fluA;ome4SUs;fluG;omeAs;rGs;fluCSup-3′ 827 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7901.26 827 omeC;omeG;omeU;rA;fluA;fluA;omeUs;ome4SG;omeAs;rGs;fluCSup-3′ 828 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7889.22 828 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;ome4SAs;rGs;fluCSup-3′ 829 5′-HO;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG; 7903.24 829 omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;ome4SGs;fluCSup-3′

Example 830

5,-ppp;rG;rC;rU;rC;rA;rU;rU;rU;rA;rC;rC;rA;rG;rC;rG;rU;rA;rA;rA;rU;rG;rA;rG;rCSu p-3′ (SEQ ID No. 839)

To fully protected CPG-bound 5′-O-dimethoxytrityl-oligoribonucleotide (44 μmol), 3% dichloroacetic acid (DCA) in methylene chloride (20 mL) was added and aged at room temperature. The DCA solution was removed after 30 min and the solid support rinsed with acetonitrile (20 mL). After removal of the acetonitrile, solutions of 5-benzylthio-1H-tetrazole (3504 μL, 0.876 μmol) and 6-chloro-N,N-diisopropyl-4H-benzo[d][1,3,2]dioxaphosphinin-2-amine (4380 μL, 0.876 μmol) in acetonitrile were added and the reaction was aged at room temperature overnight. The solution was removed from the solid support and 0.02 M I₂ in THF/H₂O/pyridine (Glen Research, cat: 40-4330-52) was added and the reaction was aged for 5 min. The oxidizer reagent was removed and the solid support was rinsed with acetonitrile (20 mL). After removal of acetonitrile, the CPG was dried under vacuum for 4 h. To the solid support was added 0.5 M tetrabutylammonium dihydrogendiphosphate in dry DMF (3.5 mL) and the mixture was aged at 45° C. for 3 days. The solid was filtered and washed with DMF, water (3×), and acetonitrile. The solid support then was treated with 8 mL AMA (40% aqueous methylamine:concentrated aqueous ammonia hydroxide, 1:1 v/v) for 2.5 h. The CPG was filtered and washed with an additional 5 mL AMA. The combined filtrates were placed in a GeneVac (ambient temperature) overnight to remove all solvent. The resulting pellet was treated with 5 mL dry THF and TBAF (tetra-n-butylammonium fluoride) in dry THF (8 mL, 1.0 M), and the mixture was aged at room temperature overnight. The reaction was quenched with 2 mL concentrated aqueous ammonia hydroxide. The solvent was removed from the reaction with a GeneVac evaporator (ambient temperature) for 5 h. The resulting viscous material was diluted with 40 mL water and dialyzed by spin filtration (3000 MWCO). The solution was further washed by spin dialysis with water (4×40 mL). The retained solution was collected, diluted with water to 15 mL and purified by ion pairing reversed phase (IP-RP) HPLC using a YMC column, 19×150 mm, 5 μm, to give the desired product.

HR LC-MS: (m/z)⁻(z=4) 1969.2855 (exp); 1969.2442 (theor).

Examples 830-837 as shown in Table 6 below were prepared according to the procedures analogous to those outlined in Example 830 above

TABLE 6 SEQ ID Ex# Sequence MW No. 830 ppp;rG;rC;rU;rC;rA; 7876.52 1 rU;rU;rU;rA;rC;rC;rA; rG;rC;rG;rU;rA;rA;rA; rU;rG;rA;rG;rCSup 831 ppp;rG;omeC,rU;rC;rA; 7956.61 1 fluU;rU;rU;rA;fluC;rC; rA;fluG;omeC;rG;fluU; omeA,fluA;omeA;rU;rG; rA;rG;omeCSup 832 ppp;rG;omeCs;rUs;rCs; 8085.13 1 rA;fluU;rU;rUs;rA; fluC;rC;rAs;fluGs; omeCs;rG;fluU;omeA; fluAs;omeA;rU;rG;rA; rG;omeCSup 833 ppp,rG;omeC;rU;rC;rA; 8036.94 1 fluU;rU;rUs;rA;fluC; rC,rAs;fluG,omeC;rG; fluU;omeA;fluA;omeA; rUs;rG;rAs;rGs;omeCSup 834 ppp;rGs;rCs;omeUs;rCs; 8177.45 1 fluAs;rUs;omeUs;rUs; rAs;rCs;omeC;rA;fluG; omeC;rG;omeU;omeA; fluA;fluA;rU;rGs; omeA,rGs;fluCSup 835 ppp;rG;rC;rU;fluC, 8097.03 1 omeA;fluU;rU;rUs;fluA; omeG;omeC;rAs;omeG; omeC,omeG;omeU;rA; fluA;fluA;omeUs;fluG, omeAs;rGs;fluCSup 836 ppp,rG;rC;rU;fluC; 8016.70 1 omeA;fluU;rU;rU;fluA; omeC;omeC;rA;omeG; omeC;omeG;omeU;rA; fluA;fluA;omeU;fluG; omeA;rG;fluCSup 837 ppp;rG;rC;rUs;fluC; 8113.09 1 omeA;fluU;rU;rUs;fluA; omeC;omeC;rAs;omeG; OmeC;omeG;omeU;rA; fluA,fluA;omeUs;fluG; omeAs;rGs;fluCSup

Example 838

5′-PcP;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;r A;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup-3′ (SEQ ID No. 840)

Fully protected CPG-bound 5′-O-dimethoxytrityl-oligoribonucleotide (5′-DMT-O;rG;rC;rUs;fluC;omeA;fluU;rU;rUs;fluA;omeC;omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA;f luA;omeUs;fluG;omeAs;rGs;fluCSup-3′ (SEQ ID NO: 842); 10 μmol) was treated with 3% 2′,2′-dichloroacetic acid in dichloromethane for 5 min (2×5 mL). Then, the solid phase was washed with acetonitrile (3×3 mL) and dichloromethane (3×3 mL). To the solid phase, N₂ was passed through for 2 min and it was kept under vacuum for 1 h.

The solid phase was treated with diethyl (((diisopropylamino)(methoxy)phosphaneyl)methyl)phosphonate (0.125 g, 0.400 mmol), acetonitrile (2.5 mL), and 5-(ethylthio)-2H-tetrazole (20 mg, 0.15 mmol) in acetonitrile (0.5 mL). The mixture was shaken at room temperature for 18 h. Then, the solid phase was filtered, rinsed with acetonitrile (2×1 mL), and treated with 12 in acetonitrile/pyridine/water (0.025 M, 9/9/2, 3 mL) for 5 min. The resin was further rinsed with acetonitrile (4×3 mL) and dichloromethane (2×3 mL). The solid phase was dried by passing a stream of N₂ through it for 1 min and kept under vacuum for 1 h.

The resulting solid phase was treated with TMSI (20 mg, 0.10 mmol) in pyridine/acetonitrile (v/v 1/50, 3 mL) for 30 min. Then, the solid phase was filtered, rinsed with acetonitrile (2×3 mL) and treated with 2-mercaptoethanol in Et₃N/acetonitrile (1 M, v/v 1/1, 0.5 mL) for 1 min. The CPG was washed again with acetonitrile (5×5 mL) and then treated with 40% aq MeNH₂ (2 mL) for 20 min. The solid support was filtered off and rinsed with 40% aq MeNH₂ (2×2 mL). The combined filtrate was shaken at ambient temperature for 2 h and then partly concentrated to ca. 2 mL under reduced pressure. The resulting solution lyophilized to give a solid.

The solid was dissolved in water (4 mL) and Et₃N-3HF (0.32 mL, 2.0 mmol) in water (1 mL) was added. The mixture was shaken at 40° C. for 6 h, after which aqueous sodium acetate (6 mL, 1 M) was added. The resulting mixture was purified by preparative HPLC (C18), eluting with 5 to 40% acetonitrile in water (0.1 M triethylammonium acetate) to provide the desired product.

LC-MS: (m/z)⁻(z=5) 1605.7 (exp), 1605.2 (theor); (z=6) 1338.0 (exp), 1337.5 (theor); (z=7) 1146.3 (exp), 1146.3 (theor); (z=8) 1002.9 (exp), 1002.9 (theor).

The oligonucleotides of the invention based on SEQ ID No. 1 comprises at least one 5′ group selected from L in Table 2a; 24 unmodified nucleobases consisting of (Adenine (A), Guanine (G), Cytosine (C) and Uracil (U), and when read from left to right, is presented as 5′-GCUCAUUUACCAGCGUAAAUGAGC-3′ (SEQ ID NO: 841) without the L group. Certain oligonucleotides of the invention based on SEQ ID No. 1 further comprise optionally modified sugar groups and optionally modified phosphodiester groups. One or more of the sugar groups optionally comprises modifications at one or more of the 2′ and/or 4′ positions selected from Table 2 for the first position nucleotide, Tables 3 and 3a for positions 2-23 nucleotides, and Table 4 for position 24 nucleotide. Exemplary modifications at the 2′ position are selected from the group consisting of OH, CH₃, OCH₃, OCF₃, OP(O)OH, O(CH₂)₂OCH₃, fluoro, NH₂, locked nucleosides, or 2′-3′ seco-nucleosides. Exemplary modifications of the 4′ position are selected from halogen, C₁₋₆ alkyl, or C₂₋₆ alkynyl. The oligonucleotides of the invention may also comprise modified phosphodiester backbone groups also depicted in Tables 3 and 3a.

An embodiment of the invention of Formula Ia and Formula I is realized when greater than 10 independently selected modified nucleobases from Table 3 are incorporated into the nucleotide sequence. A subembodiment of this aspect of the invention is realized when 10 to 20, 10 to 15, or 10 to 12 independently selected modified nucleobases from Table 3 are incorporated into the nucleotide sequence. Another subembodiment of this aspect of the invention is realized when the nucleotide sequence comprises optionally modified sugar groups and optionally modified phosphodiester backbones. A further aspect of this subembodiment of the invention is realized when one or more of the sugar groups optionally comprises modifications at one or more of the 2′ and/or 4′ positions selected from Table 2 for the first position nucleotide, Table 3 for positions 2-23 nucleotides, and Table 4 for position 24 nucleotide. Still a further aspect of this subembodiment of the invention is realized when modifications at the 2′ position are selected from the group consisting of OH, CH₃, OCH₃, OCF₃, OP(O)OH, O(CH₂)₂OCH₃, fluoro, NH₂, locked nucleosides, 2′-3′ seco-nucleosides and modifications of the 4′ position are selected from halogen, C₁₋₆ alkyl, or C₂₋₆ alkynyl.

An embodiment of the invention of Formula Ia and Formula I is realized when one or more of the sugar moieties at nucleotide positions 1, 2, 3, 7, 8, 12, and/or 17 optionally comprise a natural nucleobase; one or more of the sugar moieties at nucleotide positions 4, 6, 9, 18, 19, 21 and/or 24 optionally comprise a modified nucleobase containing a 2′ fluoro group; one or more of the sugar moieties at nucleotide positions 5, 10, 11, 13, 14, 15,16, 20, and/or 22 optionally comprise a modified nucleobase containing 2′ OMe group; and Formula Ia and Formula I optionally comprises a modified phosphodiester backbone between positions 3-4, 8-9, 20-21, 22-23 and/or 23-24.

Another embodiment of the invention of Formula Ia and Formula I is realized when SEQ ID No. 1 optionally comprises an 2′-OH at one or more of the sugar moieties at nucleotide positions 1, 3, 4, 5, 7, 8, 9, 11, 12, 15, 20, 22, and/or 23; optionally comprises fluoro at one or more of the sugar moieties at nucleotide positions 6, 10, 13, 16, and/or 18; optionally comprises 2′-OMe at one or more of the sugar moieties at nucleotide positions 2, 14, 17, 19, and/or 24; and optionally comprises a modified phosphodiester backbone between positions 8-9, 12-13, 20-21, 22-23 and/or 23-24.

An embodiment of the invention of Formula Ia and Formula I is realized when the position 8 nucleotide is selected from the group consisting of rUs, thiorUs, araU, and d5propUs.

Another embodiment of the invention of Formula Ia and Formula I is realized when the position 12 nucleotide is selected from rAs, d7dzAs, cf3As, and d2aminoAs.

Another embodiment of the invention of Formula Ia and Formula I is realized when the position 2 nucleotide is rC.

Another embodiment of the invention of Formula Ia and Formula I is realized when the position 3 nucleotide is selected from the group consisting of rUs and thiorUs.

Another embodiment of the invention of Formula Ia and Formula I is realized when the position 7 nucleotide is selected from the group consisting of rU and thiorU.

Another embodiment of the invention of Formula Ia and Formula I is realized when the position 22 nucleotide is selected from the group consisting of omeAs and araAs.

Still another embodiment of the invention of Formula Ia and Formula I is realized when the terminal 5′ substituent is selected from OH and triphosphate. A subembodiment of this aspect of the invention is realized when the terminal 5′ substituent is OH. Another subembodiment of this aspect of the invention is realized when the terminal 5′ substituent is triphosphate.

Yet another embodiment of the invention of Formula Ia and Formula I is realized when the terminal 5′ end is OH or triphosphate, nucleotide positions 1, 4, 5, 6, 9, 10, 11, 13, 14, 15, 16, 17, 18, 19, 20, 21, 23 and 24, respectively, is:

rG;rC;rUs;fluC;omeA;fluU;rU;ome4SUs;fluA;omeC;

omeC;rAs;omeG;omeC;omeG;omeU;rA;fluA;fluA;omeUs;fluG;omeAs;rGs;fluCSup (SEQ ID NO: 843). A subembodiment of this aspect of the invention is realized when nucleotide position 8 is selected from the group consisting of rUs, thiorUs, araUs, and d5propUs. Another subembodiment of this aspect of the invention is realized when nucleotide position 12 is selected from the group consisting of rAs, d7dzAs, cf3As, and d2aminoAs. Another subembodiment of this aspect of the invention is realized when nucleotide position 2 is selected from the group consisting of rC. Another subembodiment of this aspect of the invention is realized when nucleotide position 3 is selected from the group consisting of rUs and thiorUs. Another subembodiment of this aspect of the invention is realized when nucleotide position 7 is selected from the group consisting of rU and thiorU.

Biological Data

The oligonucleotides described here are RIG-I agonists which demonstrate interferon production with an EC₅₀ of <1.7 μM. Preferred oligonucleotides of the invention demonstrate EC50 values of <1.0 μM, <0.5 μM, <0.25 μM, <0.1 μM, or <0.05 μM in the A549 Dual cell assay when transfected using LyoVec. The methods below describe each of these assays.

Annealing Hairpin RNA

All hairpin RNA samples were annealed and snap cooled prior to testing. RNA samples (100 uM) were prepared in phosphate buffered saline (PBS) and transferred to a PCR plate. The RNA containing plate was sealed, placed in a thermocycler and heated at 95° C. for 1 minute followed by immediate transfer to wet ice. The plate was incubated on ice for 10 minutes and then allowed to return to room temperature prior to subsequent reformatting.

A549 Dual Cell Assay

Equilibrate a 96-well source plate(s) containing 10 nmol of lyophilized RNA in each well to room temperature. Centrifuge the plate(s) at 800×g for 5 minutes at room temperature to pellet material to the bottom of the wells. Carefully remove plate seal(s) and resuspend the RNA using 100 μL of molecular biology grade 1×PBS, hereafter referred to as 1×PBS (Thermo Fisher; Cat. No. AM9624). Mix thoroughly to ensure complete solvation of RNA. Allow the RNA plate(s) to stand for 5-10 minutes at room temperature to aid complete dissolution of all RNA. At this point the final concentration of RNA after dissolution is 100 μM in 100 μL.

To prepare dilution series of the RNA, briefly mix then transfer 60 μL of 10 μM positive control RNA (ppp-SLR10, Pyle et al. Sci. Adv. 2018 February; 4 (2): e1701854) previously prepared in 1×PBS, or test RNA solution from the 96-well source plate(s) to the appropriate wells in columns 3 and 13 of 384-well DNA Lo-Bind destination plate(s) (Eppendorf, Cat. No. 951040546). To columns 4-12 and 14-22 of the destination plate(s) add 30 μL of 1×PBS. Transfer 15 μL of RNA solution from columns 3 and 13 to adjacent wells in columns 4 and 14 and mix by aspiration. Using fresh pipette tips each time, repeat this process a further 8 times until columns 12 and 22 are reached to generate 10-point, 3-fold serial dilutions. In wells A1 to H2 and 123 to P24 add 45 μL 10 μM control RNA to define maximum effect. In wells I1 to P2 and I23 to P24 add 45 μL 1×PBS to define minimum effect. At this point these serial dilution plates can be stored at −20° C. until required.

To further dilute the RNA samples transfer 3 μL of RNA solution from all wells of the serial dilution plate(s) to fresh 384-well DNA LoBind plate(s). To these new plate(s) add 27 μL of molecular biology grade 1×PBS and mix. These plates can be stored at −20° C. until required. These 1:10 dilution daughter plates are used as the RNA source plates in this assay and can be sealed and kept at room temperature if being used on the same day or sealed and stored at −20° C. until needed.

Transfer 5 μL from each well of the 1:10 dilution daughter plate(s) to equivalent wells of 384-well assay plate(s) (Corning; Cat. No. 3764). The plates are sealed and kept at room temperature for up to 2 hours. Dissolve LyoVec transfection reagent (Invivogen; Cat. No. lyec-2) to 0.078 mg/mL using 1×PBS. Dispense 5 μL of LyoVec solution into each well of 384-well assay plate(s). Briefly centrifuge the plate(s) to ensure the RNA/LyoVec mixture is brought down to the bottom of the wells and incubate for a minimum of 15 minutes before adding cells. Rapidly thaw A549-Dual cells (originally obtained from Invivogen; Cat. No. a549d-nfisi), dilute in 5 volumes of prewarmed assay buffer (DMEM, Thermo Fisher Cat. No. 21063-029; supplemented with 10% FBS, Thermo Fisher, Cat. No. 10091148), perform a cell count on a small sample of the cell suspension and centrifuge the remainder at 200×g for 5 minutes at room temperature. Resuspend the cell pellet to a density of 2 million viable cells per mL in assay buffer. Add 20 μL of cells per well to all wells of the 384-well assay plate(s). Briefly, centrifuge the plates to ensure that the cell suspension is brought down to the bottom of the wells. Incubate the plates at 37° C., 5% CO₂ in a humidified chamber for 18 hours.

Protecting the reagent from light, reconstitute the Quanti-Luc detection reagent (Invivogen; Cat. No. rep-qlc2) with 25 mL of deionized water per reagent sachet. Remove assay plate(s) from the incubator and transfer 10 μL of supernatant from the assay plate(s) to 384-well AlphaPlate(s) (Perkin Elmer; Cat. No. 6005359). Then add 25 μL of Quanti-Luc reagent to each well of the AlphaPlate(s) and mix thoroughly. Incubate each AlphaPlate for 60 minutes at room temperature then read plate on an Envision Multilabel Plate Reader (Perkin Elmer) using a 384-L1 aperture, an integration time of 0.2 seconds and reading by rows bi-directionally.

Data is normalized to the maximum and minimum effect control wells and then subjected to a 4 parameter logistic fit based on the Levenberg-Marquardt algorithm to provide measures of potency and relative efficacy.

Examples 1 through 838 were evaluated in cell based assay as described above. Table 7 gives the biological data (RIG-I EC₅₀) collected for each example using the methodology described above.

Compounds of the disclosure were evaluated for interferon β production in A549 cell culture with LyoVec as described above. The following Table 7 tabulates the biological data for these compounds as EC₅₀.

TABLE 7 Ex EC₅₀ (μM) 1 0.002 2 0.007 3 0.012 4 0.104 5 0.197 6 0.003 7 0.069 8 0.007 9 0.006 10 0.139 11 0.007 12 0.037 13 0.029 14 0.010 15 0.049 16 0.039 17 0.016 18 0.014 19 0.061 20 0.099 21 0.009 22 0.066 23 0.176 24 0.214 15 0.058 26 0.148 27 0.016 28 0.016 29 0.018 30 0.023 31 0.005 32 0.273 33 0.005 34 0.481 35 0.029 36 0.247 37 0.048 38 0.018 39 0.008 40 0.118 41 0.169 42 0.011 43 0.172 44 0.219 45 0.038 46 0.239 47 0.066 48 0.024 49 0.101 50 0.195 51 0.080 51 0.056 53 0.350 54 0.075 55 0.022 56 0.219 57 0.011 58 0.039 59 0.375 60 0.074 61 0.121 62 0.012 63 0.008 64 0.231 65 0.006 66 0.028 67 0.036 68 0.295 69 0.084 70 0.075 71 0.422 72 0.243 73 0.250 74 0.339 75 0.194 76 0.158 77 0.038 78 0.392 79 0.121 80 0.381 81 0.310 82 0.092 83 0.280 84 0.083 85 0.011 86 0.284 87 0.012 88 0.211 89 0.295 90 0.080 91 1.122 92 0.014 93 0.011 94 0.013 95 0.013 96 0.008 97 0.012 98 0.109 99 0.018 100 0.011 101 0.014 102 0.017 103 0.128 104 1.177 105 0.104 106 0.235 107 0.240 108 0.014 109 0.021 110 0.084 111 0.014 112 0.021 113 0.073 114 0.469 115 0.292 116 0.054 117 0.137 118 0.005 119 0.026 120 0.010 121 0.029 122 0.112 123 0.139 124 0.235 125 0.322 126 1.091 127 0.253 128 0.291 129 0.077 130 0.042 131 0.120 132 0.038 133 0.368 134 0.070 135 0.114 136 1.238 137 0.160 138 0.070 139 0.065 140 0.092 141 0.319 142 0.027 143 0.272 144 0.374 145 0.055 146 0.144 147 0.121 148 0.422 149 0.013 150 0.029 151 0.551 152 0.042 153 0.022 154 0.262 155 0.377 156 0.167 157 0.200 158 0.465 159 0.065 160 0.157 161 0.143 162 0.043 163 0.384 164 0.442 165 0.246 166 0.017 167 0.442 168 0.387 169 0.076 170 0.028 171 0.538 172 0.361 173 0.206 174 0.463 175 1.101 176 0.592 177 0.502 178 0.796 179 0.002 180 0.002. 181 0.002 182 0.001 183 0.001 184 0.001 185 0.002 186 0.001 187 0.001 188 0.002 189 0.002 190 0.001 191 0.001 192 0.005 193 0.001 194 0.001 195 0.001 196 0.001 197 0.002 198 0.002 199 0.003 200 0.002 201 0.001 202 0.012 203 0.003 204 0.001 205 0.003 206 0.001 207 0.001 208 0.001 209 0.001 210 0.003 211 0.004 212 0.001 213 0.005 214 0.004 215 0.002 216 0.004 217 0.003 218 0.004 219 0.001 220 0.001 221 0.002 222 0.003 223 0.035 224 0.005 225 0.002 226 0.002. 227 0.003 228 0.005 229 0.010 230 0.003 231 0.005 232 0.018 233 0.002 234 0.004 235 0.017 236 0.004 237 0.008 238 0.006 239 0.005 240 0.001 241 0.001 242 0.041 243 0.017 244 <0.001 245 <0.001 246 0.001 247 0.001 248 0.001 249 <0.001 250 0.001 251 0.001 252 0.001 253 0.001 254 0.001 255 0.005 256 0.002 257 0.001 258 0.003 259 0.002 260 0.002 261 0.001 262 0.003 263 0.002 264 0.001 265 0.001 266 0.001 267 0.002 268 0.001 269 0.002 270 0.001 271 0.001 /72 0.002 273 0.006 274 0.004 275 0.001 276 0.012 277 0.004 278 0.005 279 0.003 280 0.003 281 0.004 282 0.013 283 0.002 284 0.002 285 0.001 286 0.001 287 0.004 288 0.012 289 0.002 290 0.003 291 0.005 292 0.002 293 0.007 294 0.006 295 0.002 296 0.006 297 0.004 298 0.003 299 0.006 300 0.002 301 0.002 302 0.002 303 0.002 304 0.002 305 0.002 306 0.006 307 0.002 308 0.023 309 0.051 310 0.004 311 0.001 312 0.003 313 0.008 314 0.004 315 0.004 316 0.864 317 0.007 318 0.040 319 0.006 320 0.016 321 0.007 322 0.010 323 0.003 324 0.009 325 0.001 326 0.005 327 0.004 328 0.012 329 0.018 330 0.015 331 0.095 332 0.009 333 0.005 334 0.003 335 <0.001 336 0.008 337 0.004 338 0.003 339 0.005 340 0.003 341 0.003 342 0.005 343 0.019 344 <0.001 345 0.003 346 0.001 347 0.006 348 0.002 349 0.002 350 0.001 351 0.002 352 0.015 353 0.006 354 0.004 355 0.004 356 0.005 357 0.003 358 0.016 359 0.004 360 0.008 361 0.028 362 0.003 363 0.003 364 0.006 365 0.037 366 0.021 367 0.055 368 0.005 369 0.012 370 0.004 371 0.001 372 <0.001 373 0.003 374 0.002 375 0.002 376 0.004 377 0.002 378 0.001 379 0.001 380 0.011 381 0.002 382 0.004 383 <0.001 384 0.005 385 0.007 386 0.005 387 0.001 388 0.005 389 0.013 390 0.020 391 0.011 392 0.015 393 0.010 394 0.003 395 0.012 396 0.005 397 0.005 398 0.010 399 0.008 400 0.006 401 0.011 402 0.007 403 0.006 404 0.011 405 0.004 406 0.005 407 0.008 408 0.004 409 0.003 410 0.005 411 0.005 412 0.006 413 0.004 414 0.005 415 0.004 416 0.005 417 0.005 418 0.004 419 0.004 420 0.003 421 0.004 422 0.006 423 0.002 424 0.006 425 0.007 426 0.010 417 0.008 428 0.004 429 0.009 430 0.005 431 0.009 432 0.003 433 0.002 434 0.004 435 0.004 436 0.010 437 0.006 438 0.006 439 0.005 440 0.008 441 0.011 442 0.007 443 0.010 444 0.012 445 0.006 446 0.004 447 0.004 448 0.001 449 0.004 450 0.003 451 0.002 452 0.002 453 0.007 454 0.029 455 0.004 456 0.003 457 0.005 458 0.004 459 0.003 460 0.004 461 0.003 462 0.004 463 0.002. 464 0.005 465 0.005 466 0.009 467 0.004 468 0.003 469 0.006 470 0.007 471 0.011 472 0.011 473 0.005 474 0.011 475 <0.001 476 0.002 477 0.004 478 0.002 479 0.008 480 0.004 481 0.004 482 0.004 483 0.004 484 0.004 485 0.004 486 0.003 487 0.003 488 0.004 489 0.003 490 0.002 491 0.004 492 0.003 493 0.002 494 0.003 495 0.002 496 0.003 497 <0.001 498 0.002 499 0.004 500 0.003 501 0.003 502 0.004 503 0.003 504 0.003 505 0.003 506 0.008 507 0.003 508 0.003 509 0.003 510 0.004 511 0.004 512 0.003 513 0.017 514 <0.001 515 0.003 516 0.002 517 0.003 518 0.001 519 0.005 520 0.010 521 0.009 522 0.026 523 0.033 524 1.111 525 1.667 526 0.009 527 0.008 528 0.008 529 0.004 530 0.005 531 0.005 532 0.004 533 0.004 534 0.003 535 0.004 536 0.002 537 0.004 538 0.022 539 0.004 540 0.004 541 0.005 542 0.004 543 0.003 544 0.004 545 0.005 546 0.004 547 0.004 548 0.004 549 0.008 550 0.005 551 0.004 552 0.011 553 0.004 554 0.004 555 0.008 556 0.382 557 0.007 558 0.010 559 0.004 560 0.004 561 0.004 562 0.004 563 0.018 564 0.013 565 0.006 566 0.007 567 0.004 568 0.006 569 0.004 570 0.011 571 0.009 572 0.004 573 0.034 574 0.008 575 0.012 576 0.004 577 0.009 578 0.012 579 0.007 580 0.010 581 0.010 582 0.007 583 0.005 584 0.005 585 0.004 586 0.008 587 0.004 588 0.012 589 0.009 590 0.009 591 0.010 592 0.013 593 0.005 594 0.014 595 0.011 596 0.012 597 0.037 598 0.040 599 0.037 600 0.029 601 0.037 602 0.018 603 <0.001 604 0.007 605 0.006 606 0.006 607 0.004 608 0.006 609 0.029 610 0.028 611 0.027 612 0.032 613 0.037 614 0.011 615 0.005 616 0.005 617 0.049 618 0.043 619 0.003 620 0.004 621 0.003 622 0.011 623 0.010 624 0.009 625 0.013 626 0.054 627 0.025 628 0.036 629 0.038 630 0.002 631 <0.001 632 0.001 633 0.001 634 0.006 635 0.011 636 0.012 637 0.036 638 0.031 639 0.032 640 0.024 641 <0.001 642 0.002 643 0.001 644 0.009 645 0.004 646 0.004 647 0.006 648 0.005 649 0.023 650 0.019 651 0.026 652 0.033 653 0.002 654 0.003 655 <0.001 656 0.002 657 0.040 658 0.042 659 0.006 660 0.002 661 0.009 662 0.007 663 0.011 664 0.011 665 0.036 666 0.037 667 0.033 668 0.038 669 <0.001 670 0.002 671 0.002 672 0.007 673 0.015 674 0.007 675 0.004 676 0.003 677 0.006 678 0.003 679 0.008 680 0.012 681 0.004 682 0.003 683 0.005 684 0.006 685 0.006 686 0.003 687 0.012 688 0.005 689 0.006 690 0.009 691 0.167 692 0.009 693 0.017 694 0.005 695 0.009 696 0.005 697 0.011 698 0.014 699 0.012 700 0.002 701 0.002 702 0.001 703 <0.001 704 0.005 705 0.013 706 0.011 707 0.006 708 0.006 709 0.008 710 0.007 711 0.010 712 0.014 713 0.011 714 0.003 715 0.003 716 0.004 717 0.004 718 0.005 719 0.007 720 0.003 721 0.008 722 0.005 723 0.003 724 0.004 725 0.003 726 0.007 727 0.003 728 0.004 729 0.004 730 0.005 731 0.007 732 0.010 733 0.006 734 0.010 735 0.111 736 0.014 737 0.142 738 0.007 739 0.013 740 0.009 741 0.009 742 0.012 743 0.007 744 0.006 745 0.001 746 0.015 747 0.012 748 0.005 749 0.010 750 0.007 751 0.011 752 0.004 753 0.009 754 0.014 755 0.012 756 0.003 757 0.007 758 0.002 759 0.005 760 0.015 761 0.011 761 0.019 763 0.009 764 0.016 765 0.007 766 0.004 767 0.009 768 0.035 769 0.007 770 0.088 771 0.019 772 0.022 773 0.062 774 0.004 775 0.002 776 0.010 777 0.003 778 0.005 779 0.008 780 0.042 781 0.009 782 0.003 783 0.003 784 0.002 785 0.006 786 0.005 787 0.007 788 0.011 789 0.011 790 0.007 791 0.023 792 0.007 793 0.010 794 0.009 795 0.006 796 0.004 797 0.009 798 0.041 799 0.017 800 0.013 801 0.004 802 0.009 803 0.006 804 0.015 805 0.009 806 0.015 807 0.009 808 0.008 809 0.013 810 0.013 811 0.013 812 0.011 813 0.011 814 0.005 815 0.008 816 0.012 817 0.007 818 0.013 819 0.003 820 0.008 821 0.006 822 0.009 823 0.004 824 0.009 825 0.009 826 0.007 827 0.016 828 0.011 829 0.025 830 <0.001 831 <0.001 832 0.001 833 <0.001 834 NA 835 NA 836 0.002 837 0.004 838 0.001 Human Peripheral Blood Mononuclear Cell (PBMC) Assay

The cell-based potencies of representative compounds of the disclosure were evaluated for interferon α production in human PBMC cell culture with LyoVec as described above compared to known literature RIG-I agonist, 5′ppp-GGACGUACGUUUCGACGUACGUCC-3′, referred to as SEQ ID No. 123 herein, and as SEQ ID No. 12 in US2016/0046943 A (5ppp 10 L); also in Linehand et al. Sci Adv. 2018 February; as 4(2) (ppp-SLR10)) were ascertained by measuring the increase of the type I interferon, IFN-α, secreted by human peripheral blood mononuclear cells (PBMCs) following transfection of the test RNAs.

The test RNAs were serially diluted in calcium/magnesium-free 1×DPBS (Gibco Cat. No. 14190136) across 10 points with 5 μL per dilution point added to the wells of a flat-bottom tissue-culture treated 384-well plate (Corning Cat. No. 3764). Five microliters per well of 1:50 diluted transfection reagent Lipofectamine 2000 (Invitrogen Cat. No. 11668019) were added to each RNA-containing. The RNA/Lipofectamine 2000 mixtures were incubated at room temperature for 10 minutes to enable complexation. Frozen human PBMCs (AllCells Cat. No. C-PB102F-001) were thawed, resuspended to a density of 2.5×10⁶/mL in assay medium (RPMI 1640 (Gibco Cat. No. 11875-085) supplemented with 10% fetal bovine serum (Gibco Cat. No. 10091148)) then 40 μL was added to each well containing the complexed RNA.

After 18 hours of incubation at 37° C., 5% CO₂, 5 μL of undiluted cell supernatant was transferred from each well into a 384-well AlphaPlate (Perkin Elmer Cat. No. 6005359). Interferon alpha levels were measured using a human IFN-α AlphaLISA Kit (Perkin Elmer Cat. No. AL297F) following the manufacturer's protocol. Briefly, 10 μL of 5× AlphaLISA anti-IFN-α acceptor beads were added to each supernatant-containing well followed by a 30 minute room temperature incubation. Ten microliters of 5× biotinylated anti-IFN-α antibody were then added to each well followed by a 60 minute room temperature incubation. Twenty-five microliters of light-sensitive 2× Streptavidin donor beads were added to each well followed by a final incubation of 30 minutes at room temperature in the dark. A Perkin Elmer Envision Multilabel Plate Reader with a 384-A1 aperture was used to scan the plates at an excitation wavelength of 680 nM and emission wavelength of 615 nM. The data were analyzed using a nonlinear regression model, log(agonist) vs. response—Variable Slope, in Prism 7.05 (GraphPad Software, La Jolla, Calif., USA). The following Table 8 tabulates the biological data for these compounds as EC₅₀.

Table 8: Biological activities of unmodified example 830 and modified example 833 compared to literature RIG-I agonist 5ppp 10 L (pppSLR10) in human PBMC assay.

TABLE 8 95% CI EC50 Lower Upper (nM) Limit Limit 5ppp 10L (pppSLR10) Donor 1 6.84 5.65 8.22 Mean EC50 (95% CI) Donor 2 7.79 5.41 10.74 nM = 6.07 (4,72, 7.75) Donor 3 4.23 3.63 4.98 Donor 4 6.04 4.48 8.21 Example 830 Donor 1 0.41 0.32 0.54 Mean EC50 (95% CI) Donor 2 0.34 0.25 0.46 nM = 0.36 (0.25, 0.51) Donor 3 0.37 0.22 0.59 Donor 4 0.32 0.22 0.46 Example 833 Donor 1 0.80 0.61 1.07 Mean EC50 (95% CI) Donor 2 0.49 0.38 0.65 nM = 0.64 (0.46, 0.93) Donor 3 0.67 0.48 0.94 Donor 4 0.63 0.39 1.14 

What is claimed is:
 1. A compound of formula Ia:

or a pharmaceutically acceptable salt thereof, wherein: each X is independently selected from the group consisting of O, S, —CH₂—, and —NH—; each Y is independently selected from the group consisting of —OH, —SH, and

each V is independently O, or S; K is H, or

each W is independently N, —CH—, or —C(F)—; each Z is independently N, or —CH—; each R₁ is independently selected from the group consisting of H, halogen, —OH, —OC₁₋₆ alkyl, C₁₋₆ alkyl, —OC₁₋₃ haloalkyl, —OC₂₋₆ alkenyl, —OC₂₋₆ alkynyl, —NH₂, and —OCH₂C₅₋₆ heteroarylCH₂R_(y), said alkyl, alkenyl, alkynyl and heteroaryl optionally substituted with 1 to 3 groups of R_(b); each R₂ is independently selected from the group consisting of H, halogen, —OH, —OC₁₋₆ alkyl, C₁₋₆ alkyl, —OC₁₋₃ haloalkyl; or one or more adjacent R₁ and R₂ can combine to form ═CH₂; each R₃ is independently H, halogen, C₁₋₆ alkyl, or C₂₋₆ alkynyl; or R₁ and R₃ on a sugar moiety can be linked to form —CH₂O, provided R₂ is hydrogen; R₄ is selected from the group consisting of —CH₂OH,

R₅ is selected from the group consisting of H, C₁₋₆ alkyl, —(CH₂CH₂O)_(p)CH₂CH₃, and —CH₂CH(OH)CH₂(O(CH₂)₂)_(n)CH₂NHC(O)R_(z); each R₆ is independently selected from H and NH₂; R_(b) is selected from C₁₋₆ alkyl, —OC₁₋₆ alkyl, halo, OH, CN, NO, NH₂, NHC₁₋₆alkyl, N(C₁₋₆alkyl)₂, SO₃H, SO₄, PO₄, C₁₋₃ haloalkyl, C₁₋₃ haloalkoxy, carboxyl, oxo, and S(O)_(g)alkyl; R is H or —P(O)(OH)_(2′); Q is independently selected from the group consisting of O, CH₂, and NH; R_(s) is NH₂ or —C(O)—; R_(x) is hydrogen or —C≡CH; R_(y) is —CH₂(O(CH₂)₂)_(n)(CH₂)_(k)NHC(O)—R_(z), or —(CH₂OCH₂)_(n)CH₂C(O)N((CH₂)_(k)NHC(O)R_(z))₂; R_(z) is cholesterol; g is 1, 2, or 3; k and n are independently selected from 1, 2, 3, 4, 5, and 6; and p is an integer from 1 to
 50. 2. A compound according to claim 1 of Formula Ia represented by Formula I:

or a pharmaceutically acceptable salt thereof, wherein: each X is independently selected from the group consisting of O, S, —CH₂—, and —NH—; each Y is independently selected from the group consisting of —OH, —SH, and

each V is independently O, or S; K is H, or

each W is independently N, —CH—, or —C(F)—; each Z is independently N, or —CH—; each R₁ is independently selected from the group consisting of H, halogen, —OH, —OC₁₋₆ alkyl, C₁₋₆ alkyl, —OC₁₋₃ haloalkyl, —OC₂₋₆ alkenyl, —OC₂₋₆ alkynyl, —NH₂, and —OCH₂C₅₋₆ heteroarylCH₂R_(y), said alkyl, alkenyl, alkynyl and heteroaryl optionally substituted with 1 to 3 groups of R_(b); each R₂ is independently selected from the group consisting of H, halogen, —OH, —OC₁₋₆ alkyl, C₁₋₆ alkyl, —OC₁₋₃ haloalkyl; or one or more adjacent R₁ and R₂ can combine to form ═CH₂; each R₃ is independently H, halogen, C₁₋₆ alkyl, or C₂₋₆ alkynyl; or R₁ and R₃ on a sugar moiety can be linked to form —CH₂O, provided R₂ is hydrogen; R₄ is selected from the group consisting of —CH₂OH,

R₅ is selected from the group consisting of H, C₁₋₆ alkyl, —(CH₂CH₂O)_(p)CH₂CH₃, and —CH₂CH(OH)CH₂(O(CH₂)₂)_(n)CH₂NHC(O)R_(z); each R₆ is independently selected from H and NH₂; R_(b) is selected from C₁₋₆ alkyl, —OC₁₋₆ alkyl, halo, OH, CN, NO, NH₂, NHC₁₋₆alkyl, N(C₁₋₆alkyl)₂, SO₃H, SO₄, PO₄, C₁₋₃ haloalkyl, C₁₋₃ haloalkoxy, carboxyl, oxo, and S(O)_(g)alkyl; R is H or —P(O)(OH)_(2′); Q is independently selected from the group consisting of O, CH₂, and NH; R_(y) is —CH₂(O(CH₂)₂)_(n)(CH₂)_(k)NHC(O)—R_(z), or —(CH₂OCH₂)_(n)CH₂C(O)N((CH₂)_(k)NHC(O)R_(z))₂; R_(z) is cholesterol; g is 1, 2, or 3; k and n are independently selected from 1, 2, 3, 4, 5, and 6; and p is an integer from 1 to
 50. 3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: any X is independently selected from O and S, any Y is independently selected from OH and SH, any V is independently selected from O and S, and any K is independently selected from H and


4. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: any R₁ is selected from the group consisting of H, halogen, —OH, —OMe, Me, —OCF₃, O(CH₂)₂OCH₃, NH₂, and —OCH₂C≡CH; any R₂ is independently selected from the group consisting of H, halogen, —OH, —OMe, Me, and —OCF₃; and Rx is hydrogen.
 5. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: any X is independently selected from O and S; any Y is independently selected from OH and SH; any V is independently selected from O and S; K is H; any W and Z within a single nine membered bicycle is independently selected from W═N and Z═—CH—, W═—CH— and Z═N and W═—CH— and Z is CH; any R₁ selected from the group consisting of H, halogen, —OH, —OMe, Me, —OCF₃, —O(CH₂)₂OCH₃, —NH₂, and —OCH₂C≡CH; any R₂ is independently selected from the group consisting of H, halogen, —OH, —OMe, -Me, and —OCF₃; R_(x) is hydrogen; R_(s) is NH₂; and any R₃ is independently selected from the group consisting of H, halogen, C₁₋₆ alkyl, and —C≡CH.
 6. A compound selected from Examples 1 through 838, or a pharmaceutically acceptable salt thereof.
 7. A pharmaceutical composition comprising a compound according to claim 6, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.
 8. The composition according to claim 7, wherein the compound comprises a hairpin or stem loop oligonucleotide further comprising a blunt end.
 9. A viral vector encoding a compound according to claim
 6. 10. An oligonucleotide capable of inducing interferon production, wherein the oligonucleotide comprises 5′-GCUCAUUUACCAGCGUAAAUGAGC-3′ (SEQ ID NO: 841), and wherein the oligonucleotide further comprises a self complementary base pairing region of less than 12 base pairs which are capable of forming a double-stranded section of the oligonucleotide.
 11. The oligonucleotide of claim 10, which forms a hairpin structure comprising the double-stranded section and a loop.
 12. The oligonucleotide of claim 10, which comprises a blunt end.
 13. The oligonucleotide of claim 10, which comprises a 5′-OH, a 5′-triphosphate, or a 5′-diphosphate.
 14. The oligonucleotide of claim 10, wherein at least one nucleotide comprises a sugar group modified at its 2′ position, wherein the 2′ modification is selected from the group consisting of: 2′-OH, 2′-methyl, 2′-OCF₃, 2′-OCH₂C≡CH, NH₂, 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyl-oxyethyl (2′-O-DMAEOE), and 2′-O—N-methylacetamido (2′-O-NMA).
 15. A pharmaceutical composition comprising an oligonucleotide of claim 10, wherein the oligonucleotide comprises at least one phosphorothioate group.
 16. The oligonucleotide of claim 10, wherein the molecule comprises at least one modified nucleobase.
 17. The compound according to claim 2, or a pharmaceutically acceptable salt thereof, wherein: any X is independently selected from O and S, any Y is independently selected from OH and SH, any V is independently selected from O and S, and any K is independently selected from H and


18. The compound according to claim 2, or a pharmaceutically acceptable salt thereof, wherein: any R₁ is selected from the group consisting of H, halogen, —OH, —OMe, Me, —OCF₃, O(CH₂)₂OCH₃, NH₂, and —OCH₂C≡CH; any R₂ is independently selected from the group consisting of H, halogen, —OH, —OMe, Me, and —OCF₃; and Rx is hydrogen.
 19. The compound according to claim 2, or a pharmaceutically acceptable salt thereof, wherein: any X is independently selected from O and S; any Y is independently selected from OH and SH; any V is independently selected from O and S; K is H; any W and Z within a single nine membered bicycle is independently selected from W═N and Z ═—CH—, W═—CH— and Z═N and W═—CH— and Z is CH; any R₁ is selected from the group consisting of H, halogen, —OH, —OMe, Me, —OCF₃, —O(CH₂)₂OCH₃, —NH₂, and —OCH₂C≡CH; any R₂ is independently selected from the group consisting of H, halogen, —OH, —OMe, -Me, and —OCF₃; R_(x) is hydrogen; R_(s) is NH₂; and any R₃ is independently selected from the group consisting of H, halogen, C₁₋₆ alkyl, and —C≡CH. 